Amorphous solid succinylated 3-(fatty acid amido)-2-hydroxy-1 -(protected hydroxy)-propane salts and methods of making the same

ABSTRACT

Aspects of the disclosure includes methods for preparing an amorphous solid composition of a fatty acid metal salt. In practicing the subject methods according to certain embodiments, a succinylated 3-(fatty acid amido)-2-hydroxy-1-(protected hydroxy)-propane organic salt is contacted with a metal base to produce a succinylated 3-(fatty acid amido)-2-hydroxy-1-(protected hydroxy)-propane metal salt; and the succinylated 3-(fatty acid amido)-2-hydroxy-1-(protected hydroxy)-propane metal salt is precipitated in a solvent to produce an amorphous solid succinylated 3-(fatty acid amido)-2-hydroxy-1-(protected hydroxy)-propane metal salt composition. An amorphous solid succinylated 3-(fatty acid amido)-2-hydroxy-1-(protected hydroxy)-propane lithium salt is also provided.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims benefit of priority to U.S. Provisional PatentApplication Ser. No. 62/926,778 filed Oct. 28, 2019, the disclosure ofwhich is herein incorporated by reference.

INTRODUCTION

Imetelstat is a telomerase inhibitor that binds with high affinity tothe template region of the RNA component of telomerase. Studies haveshown that imetelstat inhibits telomerase activity and is effectiveagainst cell proliferation in a multitude of different cancer cell linesand human tumors. Imetelstat has been used in clinical trials ofpatients with hematologic malignancies. A clinical trial of patientswith myelofibrosis showed that imetelstat was able to achieve completeclinical remissions in certain patients.

The structure of imetelstat includes a N3′→P5′ thiophosphoramidateoligonucleotide. The synthesis of imetelstat has been carried out bysolid phase oligonucleotide synthesis where the first phosphoramiditenucleotide is coupled to the support followed by sulfurization. Chainelongation of the oligonucleotide component is achieved by repeatedreaction of the 3′-amino group of the solid-phase support bondedoligonucleotide with additional nucleotide phosphoramidite monomers. Theoligonucleotide of imetelstat is coupled to the solid phase supportthrough a palmitoyl-amide linker. This fatty acid-amide linker is thus acomponent in the synthesis of imetelstat.

SUMMARY

Aspects of the disclosure includes methods for preparing an amorphoussolid composition of a fatty acid metal salt. In embodiments, thesubject methods include preparing a fatty acid metal salt from a fattyacid organic salt and precipitating the fatty acid metal salt from asolvent to produce an amorphous solid composition of the fatty acidmetal salt. In practicing the subject methods according to certainembodiments, a succinylated 3-(fatty acid amido)-2-hydroxy-1-(protectedhydroxy)-propane organic salt is contacted with a metal base to producea succinylated 3-(fatty acid amido)-2-hydroxy-1-(protectedhydroxy)-propane metal salt; and the succinylated 3-(fatty acidamido)-2-hydroxy-1-(protected hydroxy)-propane metal salt isprecipitated in a solvent to produce an amorphous solid succinylated3-(fatty acid amido)-2-hydroxy-1-(protected hydroxy)-propane metal saltcomposition. (Scheme I)

where R is a C₁₀-C₂₀ fatty acid (e.g. a C₁₆ fatty acid); and PG is aprotecting group.

In some embodiments, the metal is lithium. In some embodiments, thefatty acid is selected from palmitic acid, stearic acid, oleic acid,linoleic acid, myristoleic acid and batyl fatty acid. In someembodiments, the protected hydroxy group includes adimethoxy-triphenylmethyl protecting group. In certain embodiments, thesuccinylated 3-(fatty acid amido)-2-hydroxy-1-(protectedhydroxy)-propane metal salt is a compound of Formula I:

The amorphous composition may be formed as a crude composition. Incertain instances, the fatty acid metal salt has a purity of 95% or moreby weight of the crude composition (e.g., as determined by highperformance liquid chromatography (HPLC), proton nuclear magneticresonance spectroscopy (¹H NMR) or a combination thereof).

In some embodiments, precipitating the succinylated 3-(fatty acidamido)-2-hydroxy-1-(protected hydroxy)-propane metal salt to produce theamorphous solid composition includes: 1) heating the succinylated3-(fatty acid amido)-2-hydroxy-1-(protected hydroxy)-propane metal saltin a non-polar solvent to produce a heated succinylated 3-(fatty acidamido)-2-hydroxy-1-(protected hydroxy)-propane metal salt composition;and 2) cooling the heated succinylated 3-(fatty acidamido)-2-hydroxy-1-(protected hydroxy)-propane metal salt composition toform an amorphous solid of the succinylated 3-(fatty acidamido)-2-hydroxy-1-(protected hydroxy)-propane metal salt. In certaininstances, the succinylated 3-(fatty acid amido)-2-hydroxy-1-(protectedhydroxy)-propane metal salt is heated in the non-polar solvent to atemperature of from about 45° C. to about 55° C., such as about 50° C.To precipitate the amorphous solid composition, the heated succinylated3-(fatty acid amido)-2-hydroxy-1-(protected hydroxy)-propane metal saltcomposition may be cooled to a temperature of from about −5° C. to about5° C., such as about 0° C. to produce the amorphous solid composition.In certain instances, cooling the heated succinylated 3-(fatty acidamido)-2-hydroxy-1-(protected hydroxy)-propane metal salt compositionincludes inverse addition of the heated succinylated 3-(fatty acidamido)-2-hydroxy-1-(protected hydroxy)-propane metal salt composition toa cold non-polar solvent. The precipitated amorphous solid succinylated3-(fatty acid amido)-2-hydroxy-1-(protected hydroxy)-propane metal saltcomposition may be isolated by filtration.

In certain embodiments, methods further include preparing thesuccinylated 3-(fatty acid amido)-2-hydroxy-1-(protectedhydroxy)-propane organic salt by contacting a 3-(fatty acidamido)-2-hydroxy-1-(protected hydroxy)-propane with succinic anhydridein the presence of an organic base to succinylate the 2-hydroxy group ofthe 3-(fatty acid amido)-2-hydroxy-1-(protected hydroxy)propane. In someinstances, the 3-(fatty acid amido)-2-hydroxy-1-(protectedhydroxy)-propane is contacted with succinic anhydride in a polarsolvent.

Aspects of the disclosure also include an amorphous solid succinylated3-(fatty acid amido)-2-hydroxy-1-(protected hydroxy)-propane lithiumsalt. In some embodiments, the fatty acid is selected from palmiticacid, stearic acid, oleic acid, linoleic acid, myristoleic acid andbatyl fatty acid. In some embodiments, the protected hydroxy groupincludes a dimethoxy-triphenylmethyl protecting group. In certainembodiments, the succinylated 3-(fatty acidamido)-2-hydroxy-1-(protected hydroxy)-propane metal salt is a compoundof Formula I:

In some embodiments, the present disclosure provides an amorphous solidsuccinylated 3-(fatty acid amido)-2-hydroxy-1-(protectedhydroxy)-propane lithium salt of Formula I having a peak at about 19.5°2θ in an X-ray powder diffraction pattern (XRPD) with Cu Kα radiation.In other embodiments, the present disclosure provides an amorphous solidsuccinylated 3-(fatty acid amido)-2-hydroxy-1-(protectedhydroxy)-propane lithium salt of Formula I where thermogravimetricanalysis (TGA) exhibits a single weight loss step. In these embodiments,the single weight loss step may begin at about 225° C. The subjectamorphous solid succinylated 3-(fatty acid amido)-2-hydroxy-1-(protectedhydroxy)-propane lithium salt of Formula I may also be characterized byTGA thermogram that shows a mass loss of less than 1% of the total massof the sample upon heating from 30° C. to 300° C. In other embodiments,the present disclosure provides an amorphous solid succinylated 3-(fattyacid amido)-2-hydroxy-1-(protected hydroxy)-propane lithium salt ofFormula I, where the differential scanning calorimetry (DSC) curveexhibits an absorption peak at about 44.9° C.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 shows an X-ray powder diffraction pattern of an amorphous solidsuccinylated 3-(palmitoyl amido)-2-hydroxy-1-(dimethoxytriphenylmethylhydroxy)-propane lithium salt according to certain embodiments.

FIG. 2 shows a polarized microscopy image of an amorphous solidsuccinylated 3-(palmitoyl amido)-2-hydroxy-1-(dimethoxytriphenylmethylhydroxy)-propane lithium salt according to certain embodiments.

FIG. 3 shows a thermogram from thermogravimetric analysis (TGA) of anamorphous solid succinylated 3-(palmitoylamido)-2-hydroxy-1-(dimethoxytriphenylmethyl hydroxy)-propane lithiumsalt according to certain embodiments.

FIG. 4 shows a plot from differential scanning calorimetry of anamorphous solid succinylated 3-(palmitoylamido)-2-hydroxy-1-(dimethoxytriphenylmethyl hydroxy)-propane lithiumsalt according to certain embodiments.

SELECT DEFINITIONS OF CHEMICAL TERMINOLOGY

The following terms have the following meanings unless otherwiseindicated. Any undefined terms have their art recognized meanings.

As used herein, the terms “phosphate” and “phosphate group” are meant toencompass a thiophosphate group and an oxophosphate group.

As used herein, the term “phosphoramidite amino group” refers to theamino group, —NR⁴R⁵, attached to the phosphorus atom of aphosphoramidite group, and the term “phosphoramidite nitrogen” refers tothe nitrogen atom of the phosphoramidite amino group.

“Alkyl” refers to monovalent saturated aliphatic hydrocarbyl groupshaving from 1 to 10 carbon atoms and such as 1 to 6 carbon atoms (e.g.,“an alkyl of 1 to 6 carbons atoms”), or 1 to 5 (e.g., “an alkyl of 1 to5 carbons atoms”), or 1 to 4 (e.g., “an alkyl of 1 to 4 carbons atoms”),or 1 to 3 carbon atoms (e.g., “an alkyl of 1 to 3 carbons atoms”). Thisterm includes, by way of example, linear and branched hydrocarbyl groupssuch as methyl (CH₃—), ethyl (CH₃CH₂—), n-propyl (CH₃CH₂CH₂—), isopropyl((CH₃)₂CH—), n-butyl (CH₃CH₂CH₂CH₂—), isobutyl ((CH₃)₂CHCH₂—), sec-butyl((CH₃)(CH₃CH₂)CH—), t-butyl ((CH₃)₃C—), n-pentyl (CH₃CH₂CH₂CH₂CH₂—), andneopentyl ((CH₃)₃CCH₂—).

The term “substituted alkyl” refers to an alkyl group as defined hereinwherein one or more carbon atoms in the alkyl chain have been optionallyreplaced with a heteroatom such as —O—, —N—, —S—, —S(O)_(n)— (where n is0 to 2), —NR— (where R is hydrogen or alkyl) and having from 1 to 5substituents selected from the group consisting of alkoxy, substitutedalkoxy, cycloalkyl, substituted cycloalkyl, cycloalkenyl, substitutedcycloalkenyl, acyl, acylamino, acyloxy, amino, aminoacyl, aminoacyloxy,oxyaminoacyl, azido, cyano, halogen, hydroxyl, oxo, thioketo, carboxyl,carboxylalkyl, thioaryloxy, thioheteroaryloxy, thioheterocyclooxy,thiol, thioalkoxy, substituted thioalkoxy, aryl, aryloxy, heteroaryl,heteroaryloxy, heterocyclyl, heterocyclooxy, hydroxyamino, alkoxyamino,nitro, —SO-alkyl, —SO-aryl, —SO-heteroaryl, —SO₂-alkyl, —SO₂-aryl,—SO₂-heteroaryl, and —NR^(a)R^(b), wherein R^(a) and R^(b) may be thesame or different and are chosen from hydrogen, optionally substitutedalkyl, cycloalkyl, alkenyl, cycloalkenyl, alkynyl, aryl, heteroaryl andheterocyclic. In some instances, a“substituted alkyl” refers to an alkylgroup as defined herein having from 1 to 5 substituents selected fromthe group consisting of alkoxy, cycloalkyl, cycloalkenyl, acyl,acylamino, acyloxy, amino, aminoacyl, aminoacyloxy, oxyaminoacyl, azido,cyano, halogen, hydroxyl, carboxyl, carboxylalkyl, thiol, thioalkoxy,aryl, aryloxy, heteroaryl, heteroaryloxy, heterocyclyl, heterocyclooxy,sulfonamido, and —NR^(a)R^(b), wherein R^(a) and R^(b) may be the sameor different and are chosen from hydrogen, alkyl, cycloalkyl, alkenyl,cycloalkenyl, alkynyl, aryl, heteroaryl and heterocyclic.

“Alkylene” refers to divalent aliphatic hydrocarbyl groups preferablyhaving from 1 to 6 and more preferably 1 to 3 carbon atoms that areeither straight-chained or branched, and which are optionallyinterrupted with one or more groups selected from —O—, —NR¹⁰—,—NR¹⁰C(O)—, —C(O)NR¹⁰— and the like. This term includes, by way ofexample, methylene (—CH₂—), ethylene (—CH₂CH₂—), n-propylene(—CH₂CH₂CH₂—), iso-propylene (—CH₂CH(CH₃)—), (—C(CH₃)₂CH₂CH₂—),(—C(CH₃)₂CH₂C(O)—), (—C(CH₃)₂CH₂C(O)NH—), (—CH(CH₃)CH₂—), and the like.

“Substituted alkylene” refers to an alkylene group having from 1 to 3hydrogens replaced with substituents as described for carbons in thedefinition of “substituted” below.

The term “alkane” refers to alkyl group and alkylene group, as definedherein.

The term “alkylaminoalkyl”, “alkylaminoalkenyl” and “alkylaminoalkynyl”refers to the groups R′NHR″— where R′ is alkyl group as defined hereinand R″ is alkylene, alkenylene or alkynylene group as defined herein.

The term “alkaryl” or “aralkyl” refers to the groups -alkylene-aryl and-substituted alkylene-aryl where alkylene, substituted alkylene and arylare defined herein.

“Alkoxy” refers to the group —O-alkyl, wherein alkyl is as definedherein. Alkoxy includes, by way of example, methoxy, ethoxy, n-propoxy,isopropoxy, n-butoxy, t-butoxy, sec-butoxy, n-pentoxy, and the like. Theterm “alkoxy” also refers to the groups alkenyl-O—, cycloalkyl-O—,cycloalkenyl-O—, and alkynyl-O—, where alkenyl, cycloalkyl,cycloalkenyl, and alkynyl are as defined herein.

The term “substituted alkoxy” refers to the groups substituted alkyl-O—,substituted alkenyl-O—, substituted cycloalkyl-O—, substitutedcycloalkenyl-O—, and substituted alkynyl-O— where substituted alkyl,substituted alkenyl, substituted cycloalkyl, substituted cycloalkenyland substituted alkynyl are as defined herein.

The term “alkoxyamino” refers to the group —NH-alkoxy, wherein alkoxy isdefined herein.

The term “haloalkoxy” refers to the groups alkyl-O— wherein one or morehydrogen atoms on the alkyl group have been substituted with a halogroup and include, by way of examples, groups such as trifluoromethoxy,and the like.

The term “haloalkyl” refers to a substituted alkyl group as describedabove, wherein one or more hydrogen atoms on the alkyl group have beensubstituted with a halo group. Examples of such groups include, withoutlimitation, fluoroalkyl groups, such as trifluoromethyl, difluoromethyl,trifluoroethyl and the like.

The term “alkylalkoxy” refers to the groups -alkylene-O-alkyl,alkylene-O-substituted alkyl, substituted alkylene-O-alkyl, andsubstituted alkylene-O-substituted alkyl wherein alkyl, substitutedalkyl, alkylene and substituted alkylene are as defined herein.

The term “alkylthioalkoxy” refers to the group -alkylene-S-alkyl,alkylene-S-substituted alkyl, substituted alkylene-S-alkyl andsubstituted alkylene-S-substituted alkyl wherein alkyl, substitutedalkyl, alkylene and substituted alkylene are as defined herein.

“Alkenyl” refers to straight chain or branched hydrocarbyl groups havingfrom 2 to 6 carbon atoms and preferably 2 to 4 carbon atoms and havingat least 1 and preferably from 1 to 2 sites of double bond unsaturation.This term includes, by way of example, bi-vinyl, allyl, andbut-3-en-1-yl. Included within this term are the cis and trans isomersor mixtures of these isomers.

The term “substituted alkenyl” refers to an alkenyl group as definedherein having from 1 to 5 substituents, or from 1 to 3 substituents,selected from alkoxy, substituted alkoxy, cycloalkyl, substitutedcycloalkyl, cycloalkenyl, substituted cycloalkenyl, acyl, acylamino,acyloxy, amino, substituted amino, aminoacyl, aminoacyloxy,oxyaminoacyl, azido, cyano, halogen, hydroxyl, oxo, thioketo, carboxyl,carboxylalkyl, thioaryloxy, thioheteroaryloxy, thioheterocyclooxy,thiol, thioalkoxy, substituted thioalkoxy, aryl, aryloxy, heteroaryl,heteroaryloxy, heterocyclyl, heterocyclooxy, hydroxyamino, alkoxyamino,nitro, —SO-alkyl, —SO— substituted alkyl, —SO-aryl, —SO-heteroaryl,—SO₂-alkyl, —SO₂-substituted alkyl, —SO₂-aryl and —SO₂-heteroaryl.

“Alkynyl” refers to straight or branched monovalent hydrocarbyl groupshaving from 2 to 6 carbon atoms and preferably 2 to 3 carbon atoms andhaving at least 1 and preferably from 1 to 2 sites of triple bondunsaturation. Examples of such alkynyl groups include acetylenyl(—C≡CH), and propargyl (—CH₂C≡CH).

The term “substituted alkynyl” refers to an alkynyl group as definedherein having from 1 to 5 substituents, or from 1 to 3 substituents,selected from alkoxy, substituted alkoxy, cycloalkyl, substitutedcycloalkyl, cycloalkenyl, substituted cycloalkenyl, acyl, acylamino,acyloxy, amino, substituted amino, aminoacyl, aminoacyloxy,oxyaminoacyl, azido, cyano, halogen, hydroxyl, oxo, thioketo, carboxyl,carboxylalkyl, thioaryloxy, thioheteroaryloxy, thioheterocyclooxy,thiol, thioalkoxy, substituted thioalkoxy, aryl, aryloxy, heteroaryl,heteroaryloxy, heterocyclyl, heterocyclooxy, hydroxyamino, alkoxyamino,nitro, —SO-alkyl, —SO— substituted alkyl, —SO-aryl, —SO-heteroaryl,—SO₂-alkyl, —SO₂-substituted alkyl, —SO₂-aryl, and —SO₂-heteroaryl.

“Alkynyloxy” refers to the group —O-alkynyl, wherein alkynyl is asdefined herein. Alkynyloxy includes, by way of example, ethynyloxy,propynyloxy, and the like.

“Acyl” refers to the groups H—C(O)—, alkyl-C(O)—, substitutedalkyl-C(O)—, alkenyl-C(O)—, substituted alkenyl-C(O)—, alkynyl-C(O)—,substituted alkynyl-C(O)—, cycloalkyl-C(O)—, substitutedcycloalkyl-C(O)—, cycloalkenyl-C(O)—, substituted cycloalkenyl-C(O)—,aryl-C(O)—, substituted aryl-C(O)—, heteroaryl-C(O)—, substitutedheteroaryl-C(O)—, heterocyclyl-C(O)—, and substitutedheterocyclyl-C(O)—, wherein alkyl, substituted alkyl, alkenyl,substituted alkenyl, alkynyl, substituted alkynyl, cycloalkyl,substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl, aryl,substituted aryl, heteroaryl, substituted heteroaryl, heterocyclic, andsubstituted heterocyclic are as defined herein. For example, acylincludes the “acetyl” group CH₃C(O)—

“Acylamino” refers to the groups —NR²⁰C(O)alkyl, —NR²⁰C(O)substitutedalkyl, N R²⁰C(O)cycloalkyl, —NR²⁰C(O)substituted cycloalkyl,—NR²⁰C(O)cycloalkenyl, —NR²⁰C(O)substituted cycloalkenyl,—NR²⁰C(O)alkenyl, —NR²⁰C(O)substituted alkenyl, —NR²⁰C(O)alkynyl,—NR²⁰C(O)substituted alkynyl, —NR²⁰C(O)aryl, —NR²⁰C(O)substituted aryl,—NR²⁰C(O)heteroaryl, —NR²⁰C(O)substituted heteroaryl,—NR²⁰C(O)heterocyclic, and —NR²⁰C(O)substituted heterocyclic, whereinR²⁰ is hydrogen or alkyl and wherein alkyl, substituted alkyl, alkenyl,substituted alkenyl, alkynyl, substituted alkynyl, cycloalkyl,substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl, aryl,substituted aryl, heteroaryl, substituted heteroaryl, heterocyclic, andsubstituted heterocyclic are as defined herein.

“Aminocarbonyl” or the term “aminoacyl” refers to the group—C(O)NR²¹R²², wherein R²¹ and R²² independently are selected from thegroup consisting of hydrogen, alkyl, substituted alkyl, alkenyl,substituted alkenyl, alkynyl, substituted alkynyl, aryl, substitutedaryl, cycloalkyl, substituted cycloalkyl, cycloalkenyl, substitutedcycloalkenyl, heteroaryl, substituted heteroaryl, heterocyclic, andsubstituted heterocyclic and where R²¹ and R²² are optionally joinedtogether with the nitrogen bound thereto to form a heterocyclic orsubstituted heterocyclic group, and wherein alkyl, substituted alkyl,alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, cycloalkyl,substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl, aryl,substituted aryl, heteroaryl, substituted heteroaryl, heterocyclic, andsubstituted heterocyclic are as defined herein.

“Aminocarbonylamino” refers to the group —NR²¹C(O)NR²²R²³ where R²¹,R²², and R²³ are independently selected from hydrogen, alkyl, aryl orcycloalkyl, or where two R groups are joined to form a heterocyclylgroup.

The term “alkoxycarbonylamino” refers to the group —NRC(O)OR where eachR is independently hydrogen, alkyl, substituted alkyl, aryl, heteroaryl,or heterocyclyl wherein alkyl, substituted alkyl, aryl, heteroaryl, andheterocyclyl are as defined herein.

The term “acyloxy” refers to the groups alkyl-C(O)O—, substitutedalkyl-C(O)O—, cycloalkyl-C(O)O—, substituted cycloalkyl-C(O)O—,aryl-C(O)O—, heteroaryl-C(O)O—, and heterocyclyl-C(O)O— wherein alkyl,substituted alkyl, cycloalkyl, substituted cycloalkyl, aryl, heteroaryl,and heterocyclyl are as defined herein.

“Aminosulfonyl” refers to the group —SO₂NR²¹R²², wherein R²¹ and R²²independently are selected from the group consisting of hydrogen, alkyl,substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substitutedalkynyl, aryl, substituted aryl, cycloalkyl, substituted cycloalkyl,cycloalkenyl, substituted cycloalkenyl, heteroaryl, substitutedheteroaryl, heterocyclic, substituted heterocyclic and where R²¹ and R²²are optionally joined together with the nitrogen bound thereto to form aheterocyclic or substituted heterocyclic group and alkyl, substitutedalkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl,cycloalkyl, substituted cycloalkyl, cycloalkenyl, substitutedcycloalkenyl, aryl, substituted aryl, heteroaryl, substitutedheteroaryl, heterocyclic and substituted heterocyclic are as definedherein.

“Sulfonylamino” refers to the group —NR²¹SO₂R²², wherein R²¹ and R²²independently are selected from the group consisting of hydrogen, alkyl,substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substitutedalkynyl, aryl, substituted aryl, cycloalkyl, substituted cycloalkyl,cycloalkenyl, substituted cycloalkenyl, heteroaryl, substitutedheteroaryl, heterocyclic, and substituted heterocyclic and where R²¹ andR²² are optionally joined together with the atoms bound thereto to forma heterocyclic or substituted heterocyclic group, and wherein alkyl,substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substitutedalkynyl, cycloalkyl, substituted cycloalkyl, cycloalkenyl, substitutedcycloalkenyl, aryl, substituted aryl, heteroaryl, substitutedheteroaryl, heterocyclic, and substituted heterocyclic are as definedherein.

“Aryl” or “Ar” refers to a monovalent aromatic carbocyclic group of from6 to 18 carbon atoms having a single ring (such as is present in aphenyl group) or a ring system having multiple condensed rings (examplesof such aromatic ring systems include naphthyl, anthryl and indanyl)which condensed rings may or may not be aromatic, provided that thepoint of attachment is through an atom of an aromatic ring. This termincludes, by way of example, phenyl and naphthyl. Unless otherwiseconstrained by the definition for the aryl substituent, such aryl groupscan optionally be substituted with from 1 to 5 substituents, or from 1to 3 substituents, selected from acyloxy, hydroxy, thiol, acyl, alkyl,alkoxy, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, substituted alkyl,substituted alkoxy, substituted alkenyl, substituted alkynyl,substituted cycloalkyl, substituted cycloalkenyl, amino, substitutedamino, aminoacyl, acylamino, alkaryl, aryl, aryloxy, azido, carboxyl,carboxylalkyl, cyano, halogen, nitro, heteroaryl, heteroaryloxy,heterocyclyl, heterocyclooxy, aminoacyloxy, oxyacylamino, thioalkoxy,substituted thioalkoxy, thioaryloxy, thioheteroaryloxy, —SO-alkyl,—SO-substituted alkyl, —SO-aryl, —SO-heteroaryl, —SO₂-alkyl,—SO₂-substituted alkyl, —SO₂-aryl, —SO₂-heteroaryl and trihalomethyl. Insuch cases, an aryl group that is substituted with from 1 to 5substituents (e.g., as described herein) is referred to as a“substituted aryl”.

“Aryloxy” refers to the group —O-aryl, wherein aryl is as definedherein, including, by way of example, phenoxy, naphthoxy, and the like,including optionally substituted aryl groups as also defined herein.

“Amino” refers to the group —NH₂.

The term “substituted amino” refers to the group —NRR where each R isindependently selected from the group consisting of hydrogen, alkyl,substituted alkyl, cycloalkyl, substituted cycloalkyl, alkenyl,substituted alkenyl, cycloalkenyl, substituted cycloalkenyl, alkynyl,substituted alkynyl, aryl, heteroaryl, and heterocyclyl provided that atleast one R is not hydrogen.

The term “azido” refers to the group —N₃.

“Carboxyl,” “carboxy” or “carboxylate” refers to —CO₂H or salts thereof.

“Carboxyl ester” or “carboxy ester” or the terms “carboxyalkyl” or“carboxylalkyl” refers to the groups —C(O)O-alkyl, —C(O)O-substitutedalkyl, —C(O)O-alkenyl, —C(O)O-substituted alkenyl, —C(O)O-alkynyl,—C(O)O-substituted alkynyl, —C(O)O-aryl, —C(O)O-substituted aryl,—C(O)O-cycloalkyl, —C(O)O-substituted cycloalkyl, —C(O)O-cycloalkenyl,—C(O)O-substituted cycloalkenyl, —C(O)O-heteroaryl, —C(O)O-substitutedheteroaryl, —C(O)O-heterocyclic, and —C(O)O-substituted heterocyclic,wherein alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl,substituted alkynyl, cycloalkyl, substituted cycloalkyl, cycloalkenyl,substituted cycloalkenyl, aryl, substituted aryl, heteroaryl,substituted heteroaryl, heterocyclic, and substituted heterocyclic areas defined herein.

“(Carboxyl ester)oxy” or “carbonate” refers to the groups —O—C(O)O—alkyl, —O—C(O)O-substituted alkyl, —O—C(O)O-alkenyl,—O—C(O)O-substituted alkenyl, —O—C(O)O-alkynyl, —O—C(O)O-substitutedalkynyl, —O—C(O)O-aryl, —O—C(O)O-substituted aryl, —O—C(O)O-cycloalkyl,—O—C(O)O-substituted cycloalkyl, —O—C(O)O-cycloalkenyl, —O—C(O)O—substituted cycloalkenyl, —O—C(O)O-heteroaryl, —O—C(O)O-substitutedheteroaryl, —O—C(O)O-heterocyclic, and —O—C(O)O-substitutedheterocyclic, wherein alkyl, substituted alkyl, alkenyl, substitutedalkenyl, alkynyl, substituted alkynyl, cycloalkyl, substitutedcycloalkyl, cycloalkenyl, substituted cycloalkenyl, aryl, substitutedaryl, heteroaryl, substituted heteroaryl, heterocyclic, and substitutedheterocyclic are as defined herein.

“Cyano” or “nitrile” refers to the group —CN.

“Cycloalkyl” refers to cyclic alkyl groups of from 3 to 10 carbon atomshaving single or multiple cyclic rings including fused, bridged, andspiro ring systems. Examples of suitable cycloalkyl groups include, forinstance, adamantyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclooctyland the like. Such cycloalkyl groups include, by way of example, singlering structures such as cyclopropyl, cyclobutyl, cyclopentyl,cyclooctyl, and the like, or multiple ring structures such asadamantanyl, and the like.

The term “substituted cycloalkyl” refers to cycloalkyl groups havingfrom 1 to 5 substituents, or from 1 to 3 substituents, selected fromalkyl, substituted alkyl, alkoxy, substituted alkoxy, cycloalkyl,substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl, acyl,acylamino, acyloxy, amino, substituted amino, aminoacyl, aminoacyloxy,oxyaminoacyl, azido, cyano, halogen, hydroxyl, oxo, thioketo, carboxyl,carboxylalkyl, thioaryloxy, thioheteroaryloxy, thioheterocyclooxy,thiol, thioalkoxy, substituted thioalkoxy, aryl, aryloxy, heteroaryl,heteroaryloxy, heterocyclyl, heterocyclooxy, hydroxyamino, alkoxyamino,nitro, —SO-alkyl, —SO-substituted alkyl, —SO-aryl, —SO-heteroaryl,—SO₂-alkyl, —SO₂-substituted alkyl, —SO₂-aryl and —SO₂-heteroaryl.

“Cycloalkenyl” refers to non-aromatic cyclic alkyl groups of from 3 to10 carbon atoms having single or multiple rings and having at least onedouble bond and preferably from 1 to 2 double bonds.

The term “substituted cycloalkenyl” refers to cycloalkenyl groups havingfrom 1 to 5 substituents, or from 1 to 3 substituents, selected fromalkoxy, substituted alkoxy, cycloalkyl, substituted cycloalkyl,cycloalkenyl, substituted cycloalkenyl, acyl, acylamino, acyloxy, amino,substituted amino, aminoacyl, aminoacyloxy, oxyaminoacyl, azido, cyano,halogen, hydroxyl, keto, thioketo, carboxyl, carboxylalkyl, thioaryloxy,thioheteroaryloxy, thioheterocyclooxy, thiol, thioalkoxy, substitutedthioalkoxy, aryl, aryloxy, heteroaryl, heteroaryloxy, heterocyclyl,heterocyclooxy, hydroxyamino, alkoxyamino, nitro, —SO-alkyl,—SO-substituted alkyl, —SO-aryl, —SO-heteroaryl, —SO₂-alkyl,—SO₂-substituted alkyl, —SO₂-aryl and —SO₂-heteroaryl.

“Cycloalkynyl” refers to non-aromatic cycloalkyl groups of from 5 to 10carbon atoms having single or multiple rings and having at least onetriple bond.

“Cycloalkoxy” refers to —O-cycloalkyl.

“Cycloalkenyloxy” refers to —O-cycloalkenyl.

“Halo” or “halogen” refers to fluoro, chloro, bromo, and iodo.

“Hydroxy” or “hydroxyl” refers to the group —OH.

“Heteroaryl” refers to an aromatic group of from 1 to 15 carbon atoms,such as from 1 to 10 carbon atoms and 1 to 10 heteroatoms selected fromthe group consisting of oxygen, nitrogen, and sulfur within the ring.Such heteroaryl groups can have a single ring (such as, pyridinyl,imidazolyl or furyl) or multiple condensed rings in a ring system (forexample as in groups such as, indolizinyl, quinolinyl, benzofuran,benzimidazolyl or benzothienyl), wherein at least one ring within thering system is aromatic and at least one ring within the ring system isaromatic, provided that the point of attachment is through an atom of anaromatic ring. In certain embodiments, the nitrogen and/or sulfur ringatom(s) of the heteroaryl group are optionally oxidized to provide forthe N-oxide (N→O), sulfinyl, or sulfonyl moieties. This term includes,by way of example, pyridinyl, pyrrolyl, indolyl, thiophenyl, andfuranyl. Unless otherwise constrained by the definition for theheteroaryl substituent, such heteroaryl groups can be optionallysubstituted with 1 to 5 substituents, or from 1 to 3 substituents,selected from acyloxy, hydroxy, thiol, acyl, alkyl, alkoxy, alkenyl,alkynyl, cycloalkyl, cycloalkenyl, substituted alkyl, substitutedalkoxy, substituted alkenyl, substituted alkynyl, substitutedcycloalkyl, substituted cycloalkenyl, amino, substituted amino,aminoacyl, acylamino, alkaryl, aryl, aryloxy, azido, carboxyl,carboxylalkyl, cyano, halogen, nitro, heteroaryl, heteroaryloxy,heterocyclyl, heterocyclooxy, aminoacyloxy, oxyacylamino, thioalkoxy,substituted thioalkoxy, thioaryloxy, thioheteroaryloxy, —SO-alkyl,—SO-substituted alkyl, —SO-aryl, —SO-heteroaryl, —SO₂-alkyl,—SO₂-substituted alkyl, —SO₂-aryl and —SO₂-heteroaryl, andtrihalomethyl. In such cases, a heteroaryl group that is substitutedwith from 1 to 5 substituents (e.g., as described herein) is referred toas a “substituted heteroaryl”.

The term “heteroaralkyl” refers to the groups -alkylene-heteroaryl wherealkylene and heteroaryl are defined herein. This term includes, by wayof example, pyridylmethyl, pyridylethyl, indolylmethyl, and the like.

“Heteroaryloxy” refers to —O-heteroaryl.

“Heterocycle,” “heterocyclic,” “heterocycloalkyl,” and “heterocyclyl”refer to a saturated or unsaturated group having a single ring ormultiple condensed rings, including fused bridged and spiro ringsystems, and having from 3 to 20 ring atoms, including 1 to 10 heteroatoms. These ring atoms are selected from the group consisting ofnitrogen, sulfur, or oxygen, wherein, in fused ring systems, one or moreof the rings can be cycloalkyl, aryl, or heteroaryl, provided that thepoint of attachment is through the non-aromatic ring. In certainembodiments, the nitrogen and/or sulfur atom(s) of the heterocyclicgroup are optionally oxidized to provide for the N-oxide, —S(O)—, or—SO₂— moieties.

Examples of heterocycles and heteroaryls include, but are not limitedto, azetidine, pyrrole, imidazole, pyrazole, pyridine, pyrazine,pyrimidine, pyridazine, indolizine, isoindole, indole, dihydroindole,indazole, purine, quinolizine, isoquinoline, quinoline, phthalazine,naphthylpyridine, quinoxaline, quinazoline, cinnoline, pteridine,carbazole, carboline, phenanthridine, acridine, phenanthroline,isothiazole, phenazine, isoxazole, phenoxazine, phenothiazine,imidazolidine, imidazoline, piperidine, piperazine, indoline,phthalimide, 1,2,3,4-tetrahydroisoquinoline,4,5,6,7-tetrahydrobenzo[b]thiophene, thiazole, thiazolidine, thiophene,benzo[b]thiophene, morpholinyl, thiomorpholinyl (also referred to asthiamorpholinyl), 1,1-dioxothiomorpholinyl, piperidinyl, pyrrolidine,tetrahydrofuranyl, and the like.

Unless otherwise constrained by the definition for the heterocyclicsubstituent, such heterocyclic groups can be optionally substituted with1 to 5, or from 1 to 3 substituents, selected from alkoxy, substitutedalkoxy, cycloalkyl, substituted cycloalkyl, cycloalkenyl, substitutedcycloalkenyl, acyl, acylamino, acyloxy, amino, substituted amino,aminoacyl, aminoacyloxy, oxyaminoacyl, azido, cyano, halogen, hydroxyl,oxo, thioketo, carboxyl, carboxylalkyl, thioaryloxy, thioheteroaryloxy,thioheterocyclooxy, thiol, thioalkoxy, substituted thioalkoxy, aryl,aryloxy, heteroaryl, heteroaryloxy, heterocyclyl, heterocyclooxy,hydroxyamino, alkoxyamino, nitro, —SO-alkyl, —SO-substituted alkyl,—SO-aryl, —SO-heteroaryl, —SO₂-alkyl, —SO₂-substituted alkyl, —SO₂-aryl,—SO₂-heteroaryl, and fused heterocycle.

“Heterocyclyloxy” refers to the group —O-heterocyclyl.

The term “heterocyclylthio” refers to the group heterocyclic-S—.

The term “heterocyclene” refers to the diradical group formed from aheterocycle, as defined herein.

The term “hydroxyamino” refers to the group —NHOH.

“Nitro” refers to the group —NO₂.

“Oxo” refers to the atom (═O).

“Sulfonyl” refers to the group SO₂-alkyl, SO₂-substituted alkyl,SO₂-alkenyl, SO₂-substituted alkenyl, SO₂-cycloalkyl, SO₂-substitutedcycloalkyl, SO₂-cycloalkenyl, SO₂-substituted cycloalkenyl, SO₂-aryl,SO₂-substituted aryl, SO₂-heteroaryl, SO₂-substituted heteroaryl,SO₂-heterocyclic, and SO₂-substituted heterocyclic, wherein alkyl,substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substitutedalkynyl, cycloalkyl, substituted cycloalkyl, cycloalkenyl, substitutedcycloalkenyl, aryl, substituted aryl, heteroaryl, substitutedheteroaryl, heterocyclic, and substituted heterocyclic are as definedherein. Sulfonyl includes, by way of example, methyl-SO₂—, phenyl-SO₂—,and 4-methylphenyl-SO₂—.

“Sulfonyloxy” refers to the group —OSO₂-alkyl, OSO₂-substituted alkyl,OSO₂-alkenyl, OSO₂-substituted alkenyl, OSO₂-cycloalkyl,OSO₂-substituted cycloalkyl, OSO₂-cycloalkenyl, OSO₂-substitutedcycloalkenyl, OSO₂-aryl, OSO₂-substituted aryl, OSO₂-heteroaryl,OSO₂-substituted heteroaryl, OSO₂-heterocyclic, and OSO₂ substitutedheterocyclic, wherein alkyl, substituted alkyl, alkenyl, substitutedalkenyl, alkynyl, substituted alkynyl, cycloalkyl, substitutedcycloalkyl, cycloalkenyl, substituted cycloalkenyl, aryl, substitutedaryl, heteroaryl, substituted heteroaryl, heterocyclic, and substitutedheterocyclic are as defined herein.

The term “aminocarbonyloxy” refers to the group —OC(O)NRR where each Ris independently hydrogen, alkyl, substituted alkyl, aryl, heteroaryl,or heterocyclic wherein alkyl, substituted alkyl, aryl, heteroaryl andheterocyclic are as defined herein.

“Thiol” refers to the group —SH.

“Thioxo” or the term “thioketo” refers to the atom (═S).

“Alkylthio” or the term “thioalkoxy” refers to the group —S-alkyl,wherein alkyl is as defined herein. In certain embodiments, sulfur maybe oxidized to —S(O)—. The sulfoxide may exist as one or morestereoisomers.

The term “substituted thioalkoxy” refers to the group —S-substitutedalkyl.

The term “thioaryloxy” refers to the group aryl-S— wherein the arylgroup is as defined herein including optionally substituted aryl groupsalso defined herein.

The term “thioheteroaryloxy” refers to the group heteroaryl-S— whereinthe heteroaryl group is as defined herein including optionallysubstituted aryl groups as also defined herein.

The term “thioheterocyclooxy” refers to the group heterocyclyl-S—wherein the heterocyclyl group is as defined herein including optionallysubstituted heterocyclyl groups as also defined herein.

In addition to the disclosure herein, the term “substituted,” when usedto modify a specified group or radical, can also mean that one or morehydrogen atoms of the specified group or radical are each, independentlyof one another, replaced with the same or different substituent groupsas defined below.

In addition to the groups disclosed with respect to the individual termsherein, substituent groups for substituting for one or more hydrogens(any two hydrogens on a single carbon can be replaced with ═O, ═NR⁷⁰,═N—OR⁷⁰, ═N₂ or ═S) on saturated carbon atoms in the specified group orradical are, unless otherwise specified, —R⁶⁰, halo, ═O, —OR⁷⁰, —SR⁷⁰,—NR⁸⁰R⁸⁰, trihalomethyl, —CN, —OCN, —SCN, —NO, —NO₂, ═N₂, —N₃, —SO₂R⁷⁰,—SO₂O⁻M⁺, —SO₂OR⁷⁰, —OSO₂R⁷⁰, —OSO₂O—M⁺, —OSO₂OR⁷⁰, —P(O)(O-)₂(M⁺)₂,—P(O)(OR⁷⁰)O-M⁺, —P(O)(OR⁷⁰)₂, —C(O)R⁷⁰, —C(S)R⁷⁰, —C(NR⁷⁰)R⁷⁰,—C(O)O-M⁺, —C(O)OR⁷⁰, —C(S)OR⁷⁰, —C(O)NR⁸⁰R⁸⁰, —C(NR⁷⁰)NR⁸⁰R⁸⁰,—OC(O)R⁷⁰, —OC(S)R⁷⁰, —OC(O)O⁻M⁺, —OC(O)OR⁷⁰, —OC(S)OR⁷⁰, —NR⁷⁰C(O)R⁷⁰,—NR⁷⁰C(S)R⁷⁰, —NR⁷⁰CO₂ ⁻M⁺, —NR⁷⁰CO₂R⁷⁰, —NR⁷⁰C(S)OR⁷⁰,—NR⁷⁰C(O)NR⁸⁰R⁸⁰, —NR⁷⁰C(NR⁷⁰)R⁷⁰ and —NR⁷⁰C(NR⁷⁰)NR⁸⁰R⁸⁰, where R⁶⁰ isselected from the group consisting of optionally substituted alkyl,cycloalkyl, heteroalkyl, heterocycloalkylalkyl, cycloalkylalkyl, aryl,arylalkyl, heteroaryl and heteroarylalkyl, each R⁷⁰ is independentlyhydrogen or R⁶⁰; each R⁸⁰ is independently R⁷⁰ or alternatively, twoR⁸⁰'s, taken together with the nitrogen atom to which they are bonded,form a 5-, 6- or 7-membered heterocycloalkyl which may optionallyinclude from 1 to 4 of the same or different additional heteroatomsselected from the group consisting of O, N and S, of which N may have —Hor C₁-C₃ alkyl substitution; and each M⁺ is a counter ion with a netsingle positive charge. Each M⁺ may independently be, for example, analkali ion, such as K⁺, Na⁺, Li⁺; an ammonium ion, such as ⁺N(R⁶⁰)₄; oran alkaline earth ion, such as [Ca²⁺]_(0.5), [Mg²⁺]_(0.5), or[Ba²⁺]_(0.5) (“subscript 0.5 means that one of the counter ions for suchdivalent alkali earth ions can be an ionized form of a compound of theinvention and the other a counter ion such as chloride, or two ionizedcompounds disclosed herein can serve as counter ions for such divalentalkali earth ions, or a doubly ionized compound of the invention canserve as the counter ion for such divalent alkali earth ions). Asspecific examples, —NR⁸⁰R⁸⁰ is meant to include —NH₂, —NH-alkyl,N-pyrrolidinyl, N-piperazinyl, 4N-methyl-piperazin-1-yl andN-morpholinyl.

In addition to the disclosure herein, substituent groups for hydrogenson unsaturated carbon atoms in “substituted” alkene, alkyne, aryl andheteroaryl groups are, unless otherwise specified, —R⁶⁰, halo, —O⁻M⁺,—OR⁷⁰, —SR⁷⁰, —S⁻M⁺, —NR⁸⁰R⁸⁰, trihalomethyl, —CF₃, —CN, —OCN, —SCN,—NO, —NO₂, —N₃, —SO₂R⁷⁰, —SO₃ ⁻M⁺, —SO₃R⁷⁰, —OSO₂R⁷⁰, —OSO₃M⁺, —OSO₃R⁷⁰,—PO₃ ⁻²(M⁺)₂, —P(O)(OR⁷⁰)O⁻M⁺, —P(O)(OR⁷⁰)₂, —C(O)R⁷⁰, —C(S)R⁷⁰,—C(NR⁷⁰)R⁷⁰, —CO₂ ⁻M⁺, —CO₂R⁷⁰, —C(S)OR⁷⁰, —C(O)NR⁸⁰R⁸⁰,—C(NR⁷⁰)NR⁸⁰R⁸⁰, —OC(O)R⁷⁰, —OC(S)R⁷⁰, —OCO₂ ⁻M⁺, —OCO₂R⁷⁰, —OC(S)OR⁷⁰,—NR⁷⁰C(O)R⁷⁰, —NR⁷⁰C(S)R⁷⁰, —NR⁷⁰CO₂ ⁻M⁺, —NR⁷⁰CO₂R⁷⁰, —NR⁷⁰C(S)OR⁷⁰,—NR⁷⁰C(O)NR⁸⁰R⁸⁰, —NR⁷⁰C(NR⁷⁰)R⁷⁰ and —NR⁷⁰C(NR⁷⁰)NR⁸⁰R⁸⁰, where R⁶⁰,R⁷⁰, R⁸⁰ and M⁺ are as previously defined, provided that in case ofsubstituted alkene or alkyne, the substituents are not —O⁻M⁺, —OR⁷⁰,—SR⁷⁰, or -S⁻M⁺.

In addition to the groups disclosed with respect to the individual termsherein, substituent groups for hydrogens on nitrogen atoms in“substituted” heteroalkyl and cycloheteroalkyl groups are, unlessotherwise specified, —R⁶⁰, —O⁻M⁺, —OR⁷⁰, —SR⁷⁰, —S⁻M⁺, —NR⁸⁰R⁸⁰,trihalomethyl, —CF₃, —CN, —NO, —NO₂, —S(O)₂R⁷⁰, —S(O)₂O⁻M⁺, —S(O)₂OR⁷⁰,—OS(O)₂R⁷⁰, —OS(O)₂O⁻M⁺, —OS(O)₂OR⁷⁰, —P(O)(O⁻)₂(M⁺)₂, —P(O)(OR⁷⁰)O⁻M⁺,—P(O)(OR⁷⁰)(OR⁷⁰), —C(O)R⁷⁰, —C(S)R⁷⁰, —C(NR⁷⁰)R⁷⁰, —C(O)OR⁷⁰,—C(S)OR⁷⁰, —C(O)NR⁸⁰R⁸⁰, —C(NR⁷⁰)NR⁸⁰R⁸⁰, —OC(O)R⁷⁰, —OC(S)R⁷⁰,—OC(O)OR⁷⁰, —OC(S)OR⁷⁰, —NR⁷⁰C(O)R⁷⁰, —NR⁷⁰C(S)R⁷⁰, —NR⁷⁰C(O)OR⁷⁰,—NR⁷⁰C(S)OR⁷⁰, —NR⁷⁰C(O)NR⁸⁰R⁸⁰, —NR⁷⁰C(NR⁷⁰)R⁷⁰ and—NR⁷⁰C(NR⁷⁰)NR⁸⁰R⁸⁰, where R⁶⁰, R⁷⁰, R⁸⁰ and M⁺ are as previouslydefined.

In addition to the disclosure herein, in a certain embodiment, a groupthat is substituted has 1, 2, 3, or 4 substituents, 1, 2, or 3substituents, 1 or 2 substituents, or 1 substituent.

Unless indicated otherwise, the nomenclature of substituents that arenot explicitly defined herein are arrived at by naming the terminalportion of the functionality followed by the adjacent functionalitytoward the point of attachment. For example, the substituent“arylalkyloxycarbonyl” refers to the group (aryl)-(alkyl)-O—C(O)—.

As to any of the groups disclosed herein which contain one or moresubstituents, it is understood, of course, that such groups do notcontain any substitution or substitution patterns which are stericallyimpractical and/or synthetically non-feasible. In addition, the subjectcompounds include all stereochemical isomers arising from thesubstitution of these compounds.

“Stereoisomer” and “stereoisomers” refer to compounds that have sameatomic connectivity but different atomic arrangement in space.Stereoisomers include cis-trans isomers, E and Z isomers, enantiomers,and diastereomers.

It will be appreciated that the term “or a salt or solvate orstereoisomer thereof” is intended to include all permutations of salts,solvates and stereoisomers, such as a solvate of a pharmaceuticallyacceptable salt of a stereoisomer of subject compound. It is understoodthat the term “or a salt thereof” is intended to include allpermutations of salts. It is understood that the term “or apharmaceutically acceptable salt thereof” is intended to include allpermutations of salts. It is understood that the term “or a solvatethereof” is intended to include all permutations of solvates. It isunderstood that the term “or a stereoisomer thereof” is intended toinclude all permutations of stereoisomers. It is understood that theterm “or a tautomer thereof” is intended to include all permutations oftautomers. Thus for example it follows that it is intended to include asolvate of a pharmaceutically acceptable salt of a tautomer of astereoisomer of subject compound.

As used herein the term “isolated” is meant to describe a compound ofinterest that is in an environment different from that in which thecompound naturally occurs. “Isolated” is meant to include compounds thatare within samples that are substantially enriched for the compound ofinterest and/or in which the compound of interest is partially orsubstantially purified.

Before the present invention is further described, it is to beunderstood that this invention is not limited to particular embodimentsdescribed, as such may, of course, vary. It is also to be understoodthat the terminology used herein is for the purpose of describingparticular embodiments only, and is not intended to be limiting, sincethe scope of the present invention will be limited only by the appendedclaims.

Where a range of values is provided, it is understood that eachintervening value, to the tenth of the unit of the lower limit unlessthe context clearly dictates otherwise, between the upper and lowerlimit of that range and any other stated or intervening value in thatstated range, is encompassed within the invention. The upper and lowerlimits of these smaller ranges may independently be included in thesmaller ranges, and are also encompassed within the invention, subjectto any specifically excluded limit in the stated range. Where the statedrange includes one or both of the limits, ranges excluding either orboth of those included limits are also included in the invention.

It is appreciated that certain features of the invention, which are, forclarity, described in the context of separate embodiments, may also beprovided in combination in a single embodiment. Conversely, variousfeatures of the invention, which are, for brevity, described in thecontext of a single embodiment, may also be provided separately or inany suitable sub-combination. All combinations of the embodimentspertaining to the invention are specifically embraced by the presentinvention and are disclosed herein just as if each and every combinationwas individually and explicitly disclosed, to the extent that suchcombinations embrace subject matter that are, for example, compoundsthat are stable compounds (i.e., compounds that can be made, isolated,characterized, and tested for biological activity). In addition, allsub-combinations of the various embodiments and elements thereof (e.g.,elements of the chemical groups listed in the embodiments describingsuch variables) are also specifically embraced by the present inventionand are disclosed herein just as if each and every such sub-combinationwas individually and explicitly disclosed herein.

Unless defined otherwise, all technical and scientific terms used hereinhave the same meaning as commonly understood by one of ordinary skill inthe art to which this invention belongs. Although any methods andmaterials similar or equivalent to those described herein can also beused in the practice or testing of the present invention, methods andmaterials of interest are now described. All publications mentionedherein are incorporated herein by reference to disclose and describe themethods and/or materials in connection with which the publications arecited.

It must be noted that as used herein and in the appended claims, thesingular forms “a,” “an,” and “the” include plural referents unless thecontext clearly dictates otherwise. It is further noted that the claimsmay be drafted to exclude any optional element. As such, this statementis intended to serve as antecedent basis for use of such exclusiveterminology as “solely,” “only” and the like in connection with therecitation of claim elements, or use of a “negative” limitation.

It is appreciated that certain features of the invention, which are, forclarity, described in the context of separate embodiments, may also beprovided in combination in a single embodiment. Conversely, variousfeatures of the invention, which are, for brevity, described in thecontext of a single embodiment, may also be provided separately or inany suitable sub-combination.

The publications discussed herein are provided solely for theirdisclosure prior to the filing date of the present application. Nothingherein is to be construed as an admission that the present invention isnot entitled to antedate such publication by virtue of prior invention.Further, the dates of publication provided may be different from theactual publication dates which may need to be independently confirmed.

Except as otherwise noted, the methods and techniques of the presentembodiments are generally performed according to conventional methodswell known in the art and as described in various general and morespecific references that are cited and discussed throughout the presentspecification. See, e.g., Loudon, Organic Chemistry, Fourth Edition, NewYork: Oxford University Press, 2002, pp. 360-361, 1084-1085; Smith andMarch, March's Advanced Organic Chemistry: Reactions, Mechanisms, andStructure, Fifth Edition, Wiley-Interscience, 2001.

The nomenclature used herein to name the subject compounds isillustrated in the Examples herein. When possible, this nomenclature hasgenerally been derived using the commercially-available AutoNom software(MDL, San Leandro, Calif.).

Many general references providing commonly known chemical syntheticschemes and conditions useful for synthesizing the disclosed compoundsare available (see, e.g., Smith and March, March's Advanced OrganicChemistry: Reactions, Mechanisms, and Structure, Fifth Edition,Wiley-Interscience, 2001; or Vogel, A Textbook of Practical OrganicChemistry, Including Qualitative Organic Analysis, Fourth Edition, NewYork: Longman, 1978).

Compounds as described herein can be purified by any of the means knownin the art, including chromatographic means, such as high performanceliquid chromatography (HPLC), preparative thin layer chromatography,flash column chromatography and ion exchange chromatography. Anysuitable stationary phase can be used, including normal and reversedphases as well as ionic resins. See, e.g., Introduction to Modern LiquidChromatography, 2nd Edition, ed. L. R. Snyder and J. J. Kirkland, JohnWiley and Sons, 1979; and Thin Layer Chromatography, ed E. Stahl,Springer-Verlag, New York, 1969.

During any of the processes for preparation of the compounds of thepresent disclosure, it may be necessary and/or desirable to protectsensitive or reactive groups on any of the molecules concerned. This canbe achieved by means of conventional protecting groups as described instandard works, such as T. W. Greene and P. G. M. Wuts, “ProtectiveGroups in Organic Synthesis”, Fourth edition, Wiley, New York 2006. Theprotecting groups can be removed at a convenient subsequent stage usingmethods known from the art.

The compounds described herein can contain one or more chiral centersand/or double bonds and therefore, can exist as stereoisomers, such asdouble-bond isomers (i.e., geometric isomers), enantiomers ordiastereomers. Accordingly, all possible enantiomers and stereoisomersof the compounds including the stereoisomerically pure form (e.g.,geometrically pure, enantiomerically pure or diastereomerically pure)and enantiomeric and stereoisomeric mixtures are included in thedescription of the compounds herein. Enantiomeric and stereoisomericmixtures can be resolved into their component enantiomers orstereoisomers using separation techniques or chiral synthesis techniqueswell known to the skilled artisan. The compounds can also exist inseveral tautomeric forms including the enol form, the keto form andmixtures thereof. Accordingly, the chemical structures depicted hereinencompass all possible tautomeric forms of the illustrated compounds.The compounds described also include isotopically labeled compoundswhere one or more atoms have an atomic mass different from the atomicmass conventionally found in nature. Examples of isotopes that can beincorporated into the compounds disclosed herein include, but are notlimited to, ²H, ³H, ¹¹C, ¹³C, ¹⁴C, ¹⁵N, ¹⁸O, ¹⁷O, etc. Compounds canexist in unsolvated forms as well as solvated forms, including hydratedforms. In general, compounds can be hydrated or solvated. Certaincompounds can exist in multiple crystalline or amorphous forms. Ingeneral, all physical forms are equivalent for the uses contemplatedherein and are intended to be within the scope of the presentdisclosure.

DETAILED DESCRIPTION

As summarized above, the present disclosure provides methods forpreparing an amorphous solid composition of a fatty acid metal salt. Inpracticing the subject methods according to certain embodiments, asuccinylated 3-(fatty acid amido)-2-hydroxy-1-(protectedhydroxy)-propane organic salt is contacted with a metal base to producea succinylated 3-(fatty acid amido)-2-hydroxy-1-(protectedhydroxy)-propane metal salt; and the succinylated 3-(fatty acidamido)-2-hydroxy-1-(protected hydroxy)-propane metal salt isprecipitated in a solvent to produce an amorphous solid composition ofthe fatty acid metal salt. The term “succinylated” is used herein in itsconventional sense to refer to addition of a succinyl group(—CO—CH₂—CH₂—CO₂H) to the 3-(fatty acid amido)-2-hydroxy-1-(protectedhydroxy)-propane (e.g., at the C-2 position of the 3-(fatty acidamido)-2-hydroxy-1-(protected hydroxy)-propane). In embodiments, thesuccinyl group can be added to the 3-(fatty acidamido)-2-hydroxy-1-(protected hydroxy)-propane by any convenientsuccinylation protocol, such as for example by reaction with succinicanhydride in the presence of a base (e.g., organic base, such astriethylamine). By “fatty acid amido” is meant a structural moiety thatincludes an aliphatic group bonded to an amide group (—R—CO—N—R′—, whereR is an aliphatic chain of C₁₀-C₂₀, such as C₁₆ and R′ is the C3 carbonof the 2-hydroxy-1-(protected hydroxy) propane). In embodiments (asdescribed in greater detail below), the fatty acid may be an aliphaticchain C₁₀-C₂₀, such as C₁₆ alkyl chain. The fatty acid component may besaturated or may include one or more unsaturations. In some instances,the fatty acid component is fully saturated. In other instances, thefatty acid component is monounsaturated. In yet other instances, thefatty acid component is polyunsaturated, such as having 2, 3, 4 or moresaturations. In some embodiments, the fatty acid is selected frompalmitic acid, stearic acid, oleic acid, linoleic acid, myristoleic acidand batyl fatty acid.

Embodiments of the present disclosure describe an amorphous solidcomposition of a fatty acid metal salt. The term “amorphous” is usedherein in its conventional sense to refer to a solid material that ischaracterized by undefined structural order and microscopicconfigurations that lack a regular geometric arrangement in threedimensions. In some embodiments, the amorphous solid composition of thefatty acid metal salt are not crystalline (i.e., a solid material wherethe molecules that form the solid are arranged in a highly orderedmicroscopic geometric configuration (e.g., form an ordered lattice-typestructure) that extends in three dimensions)

In embodiments, the organic salt of the succinylated 3-(fatty acidamido)-2-hydroxy-1-(protected hydroxy)-propane may include, but is notlimited to, a triethylamine salt, triethanolamine salt, an ammoniumsalt, an arginine salt, a benzathine salt, an ethylenediamine salt, ameglumine salt, a procaine salt, an N-methylglucamine salt, a piperazinesalt, a tromethamine salt, an N,N′-dibenzylethylene-diamine salt, achloroprocaine salt, a diethanolamine salt, an ethanolamine salt, adiisopropylamine salt, a diisopropylethylamine salt, among other organiccation salts. In certain embodiments, the succinylated 3-(fatty acidamido)-2-hydroxy-1-(protected hydroxy)-propane organic salt is asuccinylated 3-(fatty acid amido)-2-hydroxy-1-(protectedhydroxy)-propane triethylamine salt.

The succinylated 3-(fatty acid amido)-2-hydroxy-1-(protectedhydroxy)-propane organic salt is contacted with a metal base in asolvent. Depending on the desired succinylated 3-(fatty acidamido)-2-hydroxy-1-(protected hydroxy)-propane metal salt, the metalbase may include, but is not limited to lithium tert-butoxide, lithiumhydroxide, lithium ethoxide, lithium isopropoxide, lithium methoxide,potassium tert-butoxide, potassium hydroxide, potassium ethoxide,potassium isopropoxide, potassium methoxide, sodium tert-butoxide,sodium hydroxide, sodium ethoxide, sodium isopropoxide, sodiummethoxide, magnesium tert-butoxide, magnesium hydroxide, magnesiumethoxide, magnesium isopropoxide, magnesium methoxide, calciumtert-butoxide, calcium hydroxide, calcium ethoxide, calciumisopropoxide, calcium methoxide. In embodiments, the succinylated3-(fatty acid amido)-2-hydroxy-1-(protected hydroxy)-propane organicsalt is contacted in an amount sufficient to generate a succinylated3-(fatty acid amido)-2-hydroxy-1-(protected hydroxy)-propane metal saltsuch as a succinylated 3-(fatty acid amido)-2-hydroxy-1-(protectedhydroxy)-propane lithium salt, a succinylated 3-(fatty acidamido)-2-hydroxy-1-(protected hydroxy)-propane sodium salt, asuccinylated 3-(fatty acid amido)-2-hydroxy-1-(protectedhydroxy)-propane potassium salt, a succinylated 3-(fatty acidamido)-2-hydroxy-1-(protected hydroxy)-propane magnesium salt or asuccinylated 3-(fatty acid amido)-2-hydroxy-1-(protectedhydroxy)-propane calcium salt.

The amount of metal base contacted with the succinylated 3-(fatty acidamido)-2-hydroxy-1-(protected hydroxy)-propane organic salt may vary,ranging from 0.5 equivalents to 2 equivalents of metal base tosuccinylated 3-(fatty acid amido)-2-hydroxy-1-(protectedhydroxy)-propane organic salt, such as from 0.75 equivalents to 1.85equivalents, such as 1 equivalent to 1.8 equivalents, such as from 1.05equivalents to 1.75 equivalents, such as from 1.10 equivalents to 1.70equivalents, such as from 1.15 equivalents to 1.65 equivalents, such asfrom 1.2 equivalents to 1.6 equivalents, such as from 1.25 equivalentsto 1.55 equivalents and including from 1.3 equivalents to 1.5equivalents and including contacting the succinylated 3-(fatty acidamido)-2-hydroxy-1-(protected hydroxy)-propane organic salt with 1.5equivalents of metal base.

In embodiments, the succinylated 3-(fatty acidamido)-2-hydroxy-1-(protected hydroxy)-propane organic salt is contactedwith the metal base in a solvent. The solvent for contacting thesuccinylated 3-(fatty acid amido)-2-hydroxy-1-(protectedhydroxy)-propane organic salt with the metal base may be any suitablesolvent where the succinylated 3-(fatty acidamido)-2-hydroxy-1-(protected hydroxy)-propane organic salt and metalbase are sufficiently soluble and may be polar or nonpolar. In someembodiments, the solvent is non-polar, such as pentane, hexane, heptane,octane or benzene. In other embodiments, the solvent is a polar solvent,such as dichloromethane, tetrahydrofuran, methyltetrahydrofuran,isopropylacetate, dimethylformamide, acetonitrile, toluene,2-methylbutan-2-ol (tAmOH) and N-methyl-2-pyrrolidone. In certainembodiments, the succinylated 3-(fatty acidamido)-2-hydroxy-1-(protected hydroxy)-propane organic salt is contactedwith the metal base in dichloromethane. In some embodiments, thesuccinylated 3-(fatty acid amido)-2-hydroxy-1-(protectedhydroxy)-propane organic salt is contacted with the metal base bycontacting the succinylated 3-(fatty acid amido)-2-hydroxy-1-(protectedhydroxy)-propane organic salt with a solvent containing the metal base.In these embodiments, the solvent containing the metal base may be anon-polar solvent, such as pentane, hexane, heptane, octane or benzene.In certain embodiments, the solvent is hexane.

The succinylated 3-(fatty acid amido)-2-hydroxy-1-(protectedhydroxy)-propane organic salt may be contacted with the metal base at atemperature that ranges from −10° C. to 10° C., such as from −9° C. to9° C., such as from −8° C. to 8° C., such as from −7° C. to 7° C., suchas from −6° C. to 6° C. and including from −5° C. to 5° C.

In some embodiments, the succinylated 3-(fatty acidamido)-2-hydroxy-1-(protected hydroxy)-propane organic salt may becontacted with the metal base at a first temperature and the reactionmixture is warmed to a second temperature. In one example, thesuccinylated 3-(fatty acid amido)-2-hydroxy-1-(protectedhydroxy)-propane organic salt is contacted with the metal base in thesolvent at a first temperature and then warmed to a second temperature.In another example, the succinylated 3-(fatty acidamido)-2-hydroxy-1-(protected hydroxy)-propane organic salt is contactedwith the metal base in the solvent at a first temperature for a firstperiod of time and then warmed to a second temperature for a secondperiod of time. In these embodiments, the first temperature may vary,ranging from −10° C. to 0° C., such as from −9° C. to −1° C., such asfrom −8° C. to −2° C., such as from −7° C. to −3° C. and including from−6° C. to −4° C. The second temperature may also range, such as from 0°C. to 20° C., such as from 1° C. to 19° C., such as from 2° C. to 18°C., such as from 3° C. to 17° C., such as from 4° C. to 16° C., such asfrom 5° C. to 15° C. and including from 5° C. to 10° C. The duration ofthe first period of time may vary depending on the concentration of themetal salt and temperature and may range from 0.1 minutes to 30 minutes,such as from 0.5 minutes to 25 minutes, such as from 1 minute to 20minutes, such as from 5 minutes to 15 minutes and including from 5minutes to 10 minutes. In some embodiments, the metal base is contactedwith the succinylated 3-(fatty acid amido)-2-hydroxy-1-(protectedhydroxy)-propane organic salt and maintained in contact (e.g., stirringthe reaction mixture) for the entire duration of the first period oftime. In other embodiments, the metal base is dropwise added (ortitrated) over all or part of the first period of time to thesuccinylated 3-(fatty acid amido)-2-hydroxy-1-(protectedhydroxy)-propane organic salt in the solvent. The duration of the secondperiod of time may vary, ranging from 0.1 minutes to 30 minutes, such asfrom 0.5 minutes to 25 minutes, such as from 1 minute to 20 minutes,such as from 5 minutes to 15 minutes and including from 5 minutes to 10minutes.

In some embodiments, methods include precipitating the succinylated3-(fatty acid amido)-2-hydroxy-1-(protected hydroxy)-propane metal saltfrom the reaction mixture to form an amorphous solid. To precipitate theamorphous solid succinylated 3-(fatty acid amido)-2-hydroxy-1-(protectedhydroxy)-propane metal salt, the reaction mixture is contacted with thesame or another nonpolar solvent to generate an amorphous solidsuccinylated 3-(fatty acid amido)-2-hydroxy-1-(protectedhydroxy)-propane metal salt composition. The nonpolar solvent mayinclude, but is not limited to pentane, hexanes, heptane, octane,benzene, among other nonpolar solvents. In certain embodiments, thesuccinylated 3-(fatty acid amido)-2-hydroxy-1-(protectedhydroxy)-propane metal salt reaction mixture is contacted with heptane.

In some embodiments, methods include first cooling the succinylated3-(fatty acid amido)-2-hydroxy-1-(protected hydroxy)-propane metal saltreaction mixture before contacting the reaction mixture with thenon-polar solvent (e.g., heptane). For example, the reaction mixture maybe cooled to a temperature that ranges from −10° C. to 10° C., such asfrom −9° C. to 9° C., such as from −8° C. to 8° C., such as from −7° C.to 7° C., such as from −6° C. to 6° C. and including from −5° C. to 5°C.

In precipitating the amorphous solid succinylated 3-(fatty acidamido)-2-hydroxy-1-(protected hydroxy)-propane metal salt, methodsaccording to certain embodiments include concentrating the succinylated3-(fatty acid amido)-2-hydroxy-1-(protected hydroxy)-propane metal saltin the non polar solvent (e.g., heptane), such as by heating orrotoevaporation. The volume of solvent in the the succinylated 3-(fattyacid amido)-2-hydroxy-1-(protected hydroxy)-propane reaction mixture maybe reduced by 5% or more, such as by 10% or more, such as by 15% ormore, such as by 20% or more, such as by 25% or more, such as by 30% ormore, such as by 35% or more, such as by 40% or more, such as by 45% ormore, such as by 50% or more, such as by 55% or more, such as by 60% ormore, such as by 65% or more, such as 70% or more and including by 75%or more. This may be repeated one or more times, where an amount of thenon-polar solvent is added to the succinylated 3-(fatty acidamido)-2-hydroxy-1-(protected hydroxy)-propane metal salt compositionand then further concentrated, such as repeated two or more times, suchas three or more times and including five or more times.

To precipitate the amorphous succinylated 3-(fatty acidamido)-2-hydroxy-1-(protected hydroxy)-propane metal salt from thenon-polar solvent, the composition in the non-polar solvent (e.g.,heptane) may be first heated to produce a heated succinylated 3-(fattyacid amido)-2-hydroxy-1-(protected hydroxy)-propane metal saltcomposition and then cooled to form the amorphous solid succinylated3-(fatty acid amido)-2-hydroxy-1-(protected hydroxy)-propane metal salt.The succinylated 3-(fatty acid amido)-2-hydroxy-1-(protectedhydroxy)-propane metal salt may be heated in the nonpolar solvent to atemperature that ranges from 20° C. to 60° C., such as from 25° C. to60° C., such as from 40° C. to 55° C. and including from 45° C. to 55°C. The heated composition may be maintained at the elevated temperaturefor a duration that varies, such as for 1 minute or longer, such as for2 minutes or longer, such as for 5 minutes or longer, such as 10 minutesor longer, such as 15 minutes or longer, such as 30 minutes or longerand including for 60 minutes or longer. In certain embodiments, methodsinclude heating the composition until the succinylated 3-(fatty acidamido)-2-hydroxy-1-(protected hydroxy)-propane metal salt is solubilizedinto the solvent, such as by observing a clear solution by visualinspection.

The heated composition is cooled to precipitate the succinylated3-(fatty acid amido)-2-hydroxy-1-(protected hydroxy)-propane metal salt.The heated composition is cooled to a temperature of −20° C. to 20° C.,such as from −19° C. to 19° C., such as from −18° C. to 18° C., such asfrom −17° C. to 17° C., such as from −16° C. to 16° C., such as from−15° C. to 15° C., such as from −14° C. to 14° C., such as from −13° C.to 13° C. such as from −12° C. to 12° C. such as from −11° C. to 11° C.,such as from −10° C. to 10° C., and including from −5° C. to 5° C. Incertain embodiments, the heated succinylated 3-(fatty acidamido)-2-hydroxy-1-(protected hydroxy)-propane metal salt composition iscooled to 0° C. The cooled composition may be maintained at the reducedtemperature (e.g., at 0° C.) for a duration that varies, such as for 1minute or longer, such as for 2 minutes or longer, such as for 5 minutesor longer, such as 10 minutes or longer, such as 15 minutes or longer,such as 30 minutes or longer and including for 60 minutes or longer. Incertain embodiments, the cooled succinylated 3-(fatty acidamido)-2-hydroxy-1-(protected hydroxy)-propane metal salt composition ismaintained at the reduced temperature (e.g., at 0° C.) for 60 minutes.

The amorphous solid succinylated 3-(fatty acidamido)-2-hydroxy-1-(protected hydroxy)-propane metal salt compositionmay be isolated by filtration (e.g., vacuum filtration) or the solventmay be removed by heating or rotoevaporation. In certain embodiments,the amorphous solid succinylated 3-(fatty acidamido)-2-hydroxy-1-(protected hydroxy)-propane metal salt composition isisolated by drying at room temperature under nitrogen atmosphere orunder vacuum. In some embodiments, the amorphous solid succinylated3-(fatty acid amido)-2-hydroxy-1-(protected hydroxy)-propane metal saltis isolated at a temperature of −20° C. to 20° C., such as from −19° C.to 19° C., such as from −18° C. to 18° C., such as from −17° C. to 17°C., such as from −16° C. to 16° C., such as from −15° C. to 15° C., suchas from −14° C. to 14° C., such as from −13° C. to 13° C. such as from−12° C. to 12° C. such as from −11° C. to 11° C., such as from −10° C.to 10° C. and including from −5° C. to 5° C. In certain embodiments, theamorphous solid succinylated 3-(fatty acid amido)-2-hydroxy-1-(protectedhydroxy)-propane metal salt is isolated at 0° C.

In some embodiments, methods include precipitating the amorphous solidsuccinylated 3-(fatty acid amido)-2-hydroxy-1-(protectedhydroxy)-propane metal salt by an inverse addition of a non-polarsolvent to the succinylated 3-(fatty acid amido)-2-hydroxy-1-(protectedhydroxy)-propane metal salt reaction mixture. In this embodiment, anamount of the succinylated 3-(fatty acid amido)-2-hydroxy-1-(protectedhydroxy)-propane metal salt reaction mixture is contacted with anon-polar solvent. The non-polar solvent may include, but is not limitedto pentane, hexanes, heptane, octane, benzene, among other nonpolarsolvents. In certain instances, the succinylated 3-(fatty acidamido)-2-hydroxy-1-(protected hydroxy)-propane metal salt reactionmixture is contacted with heptane.

The reaction mixture contacted with the non-polar solvent isconcentrated, such as by heating or rotoevaporation. The volume of thereaction mixture may be reduced by 5% or more, such as by 10% or more,such as by 15% or more, such as by 20% or more, such as by 25% or more,such as by 30% or more, such as by 35% or more, such as by 40% or more,such as by 45% or more, such as by 50% or more, such as by 55% or more,such as by 60% or more, such as by 65% or more, such as 70% or more andincluding by 75% or more. This may be repeated one or more times, wherean amount of the non-polar solvent is added to the succinylated 3-(fattyacid amido)-2-hydroxy-1-(protected hydroxy)-propane metal saltcomposition and then further concentrated, such as repeated two or moretimes, such as three or more times and including five or more times.

To precipitate the amorphous succinylated 3-(fatty acidamido)-2-hydroxy-1-(protected hydroxy)-propane metal salt from thenon-polar solvent, the composition is heated to produce a heatedsuccinylated 3-(fatty acid amido)-2-hydroxy-1-(protectedhydroxy)-propane metal salt composition. The composition may be heatedin the nonpolar solvent to a temperature that ranges from 20° C. to 60°C., such as from 25° C. to 60° C., such as from 40° C. to 55° C. andincluding from 45° C. to 55° C. The heated composition may be maintainedat the elevated temperature for a duration that varies, such as for 1minute or longer, such as for 2 minutes or longer, such as for 5 minutesor longer, such as 10 minutes or longer, such as 15 minutes or longer,such as 30 minutes or longer and including for 60 minutes or longer. Incertain embodiments, methods include heating the composition until thesuccinylated 3-(fatty acid amido)-2-hydroxy-1-(protectedhydroxy)-propane metal salt is solubilized into the solvent, such as byobserving a clear solution by visual inspection.

To cool the heated succinylated 3-(fatty acidamido)-2-hydroxy-1-(protected hydroxy)-propane metal salt composition,the heated composition is contacted with a cold non-polar solvent (e.g.,heptane) In these embodiments, the heated succinylated 3-(fatty acidamido)-2-hydroxy-1-(protected hydroxy)-propane metal salt compositionmay be added to a non-polar solvent (e.g., heptane) that is at atemperature of −20° C. to 20° C., such as from −19° C. to 19° C., suchas from −18° C. to 18° C., such as from −17° C. to 17° C., such as from−16° C. to 16° C., such as from −15° C. to 15° C., such as from −14° C.to 14° C., such as from −13° C. to 13° C. such as from −12° C. to 12° C.such as from −11° C. to 11° C., such as from −10° C. to 10° C. andincluding from −5° C. to 5° C. In certain embodiments, the heatedsuccinylated 3-(fatty acid amido)-2-hydroxy-1-(protectedhydroxy)-propane metal salt composition is contacted with a non-polarsolvent that is at a temperature of −10° C. The cooled composition maybe maintained at the reduced temperature (e.g., at 0° C.) for a durationthat varies, such as for 0.1 hours or longer, such as for 0.5 hours orlonger, such as for 1 hour or longer, such as for 1.5 hours or longer,such as for 2 hours or longer, such as for 2.5 hours or longer andincluding for 3 hours or longer. In certain embodiments, the cooledsuccinylated 3-(fatty acid amido)-2-hydroxy-1-(protectedhydroxy)-propane metal salt composition is maintained at the reducedtemperature (e.g., at 0° C.) for 3 hours.

In certain embodiments, methods include conducting one or moreheating/cooling cycles on the succinylated 3-(fatty acidamido)-2-hydroxy-1-(protected hydroxy)-propane metal salt composition.In these embodiments, each heating/cooling cycle includes heating thesuccinylated 3-(fatty acid amido)-2-hydroxy-1-(protectedhydroxy)-propane metal salt composition to a first temperature andmaintaining the composition at the first temperature for a first periodof time, followed by cooling the heated composition to a secondtemperature and maintaining the cooled compositioned at the secondtemperature for a second period of time. In these embodiments, the firsttemperature ranges from 10° C. to 30° C., such as from 15° C. to 25° C.and including to a temperature of 20° C. and the first period of timeranges from 30 minutes to 60 minutes, such as from 35 minutes to 55minutes and including from 40 minutes to 50 minutes. The secondtemperature ranges from −10° C. to 10° C., such as from −5° C. to 5° C.and including 0° C. and the second period of time ranges from 45 minutesto 90 minutes, such as from 50 minutes to 85 minutes, such as from 55minutes to 80 minutes and including from 60 minutes to 75 minutes. Theheating/cooling cycle may be repeated one or more times, such as two ormore times and including three or more times.

The precipitated succinylated 3-(fatty acidamido)-2-hydroxy-1-(protected hydroxy)-propane metal salt composition inthe non-polar solvent may be maintained at a reduced temperature for aperiod of time before isolation. In some instances, the precipitatedsuccinylated 3-(fatty acid amido)-2-hydroxy-1-(protectedhydroxy)-propane metal salt composition is maintained in the non-polarsolvent at a temperature ranges from −10° C. to 10° C., such as from −5°C. to 5° C. and including 0° C. The precipitated succinylated 3-(fattyacid amido)-2-hydroxy-1-(protected hydroxy)-propane metal saltcomposition may be maintained at the reduced temperature for a period oftime that varies, such as for 1 hour or longer, such as for 2 hours orlonger, such as for 4 hours or longer, such as for 8 hours or longer,such as from 12 hours or longer and including for 16 hours or longer.

The amorphous solid succinylated 3-(fatty acidamido)-2-hydroxy-1-(protected hydroxy)-propane metal salt compositionmay be isolated by filtration (e.g., vacuum filtration) or the solventmay be removed by heating or rotoevaporation. In certain embodiments,the amorphous solid succinylated 3-(fatty acidamido)-2-hydroxy-1-(protected hydroxy)-propane metal salt composition isisolated by drying at room temperature under nitrogen atmosphere orunder vacuum. In some embodiments, the amorphous solid succinylated3-(fatty acid amido)-2-hydroxy-1-(protected hydroxy)-propane metal saltis isolated at a temperature of −20° C. to 20° C., such as from −19° C.to 19° C., such as from −18° C. to 18° C., such as from −17° C. to 17°C., such as from −16° C. to 16° C., such as from −15° C. to 15° C., suchas from −14° C. to 14° C., such as from −13° C. to 13° C. such as from−12° C. to 12° C. such as from −11° C. to 11° C., such as from −10° C.to 10° C. and including from −5° C. to 5° C. In certain embodiments, theamorphous solid succinylated 3-(fatty acid amido)-2-hydroxy-1-(protectedhydroxy)-propane metal salt is isolated at 0° C. The isolated amorphoussolid succinylated 3-(fatty acid amido)-2-hydroxy-1-(protectedhydroxy)-propane metal salt may be dried to remove any residual solvent,such as under vacuum at ambient temperature.

In certain embodiments, methods further include preparing thesuccinylated 3-(fatty acid amido)-2-hydroxy-1-(protectedhydroxy)-propane organic salt by contacting a 3-(fatty acidamido)-2-hydroxy-1-(protected hydroxy)-propane with succinic anhydridein the presence of an organic base. The amount of succinic anhydridecontacted with the 3-(fatty acid amido)-2-hydroxy-1-(protectedhydroxy)-propane may vary, ranging from 0.5 equivalents to 2 equivalentsof succinic anhydride to 3-(fatty acid amido)-2-hydroxy-1-(protectedhydroxy)-propane, such as from 0.75 equivalents to 1.95 equivalents,such as 1 equivalent to 1.9 equivalents, such as from 1.1 equivalents to1.85 equivalents, such as from 1.15 equivalents to 1.80 equivalents,such as from 1.25 equivalents to 1.75 equivalents and includingcontacting the 3-(fatty acid amido)-2-hydroxy-1-(protectedhydroxy)-propane with 1.2 equivalents of succinic anhydride.

Depending on the type of organic salt desired, the organic base used mayinclude, but is not limited to, triethylamine, triethanolamine, ammonia,arginine, benzathine, ethylenediamine, meglumine, procaine,N-methylglucamine, piperazine, tromethamine,N,N′-dibenzylethylene-diamine, chloroprocaine, diethanolamine,ethanolamine, diisopropylamine, diisopropylethylamine, among otherorganic bases. In these embodiments, the 3-(fatty acidamido)-2-hydroxy-1-(protected hydroxy)-propane is contacted withsuccinic anhydride in a polar solvent, such as dichloromethane,tetrahydrofuran, methyltetrahydrofuran, isopropylacetate,dimethylformamide, acetonitrile, toluene, 2-methylbutan-2-ol (tAmOH) andN-methyl-2-pyrrolidone. The amount of organic base used may vary,ranging from 2.5 equivalents to 3.5 equivalents of organic base to3-(fatty acid amido)-2-hydroxy-1-(protected hydroxy)-propane, such asfrom 2.6 equivalents to 3.4 equivalents, such as 2.7 equivalent to 3.3equivalents, such as from 2.8 equivalents to 3.2 equivalents, such asfrom 2.9 equivalents to 3.1 equivalents and including contacting the3-(fatty acid amido)-2-hydroxy-1-(protected hydroxy)-propane with 3.0equivalents of organic base (e.g., trimethylamine).

The 3-(fatty acid amido)-2-hydroxy-1-(protected hydroxy)-propane may becontacted with succinic anhydride in the presence of the organic base ata temperature that ranges from 10° C. to 30° C., such as from 12° C. to28° C., such as from 14° C. to 26° C., such as from 16° C. to 24° C. andincluding from 18° C. to 22° C.

The components used in each step of the subject methods for preparingthe amorphous succinylated 3-(fatty acid amido)-2-hydroxy-1-(protectedhydroxy)-propane metal salt described herein may be a purifiedcomposition or a crude composition as desired. The term “purified” isused in its conventional sense to refer to a composition where at leastsome isolation or purification process has been conducted, such as forexample, filtration or aqueous workup of a reaction mixture. In certaininstances, purification includes liquid chromatography,recrystallization, distillation (e.g., azeotropic distillation) or othertype of compound purification. In some embodiments, a reaction mixtureis used in a subsequent step in the methods described herein as a crudemixture. The term “crude” is used herein in its conventional sense torefer to a composition where no purification or other workup of thereaction mixture has been conducted. In certain instances, the crudecomposition reaction mixtures include the compound of interest insufficient purity such as where the crude composition includes acompound of interest in a purity of 90% or greater, 91% or greater, 92%or greater, 93% or greater, 94% or greater, such as 95% or greater, 96%or greater, such as 97% or greater, 98% or greater, and including 99% orgreater, as determined by high performance liquid chromatography (HPLC),proton nuclear magnetic resonance spectroscopy (¹H NMR) or a combinationthereof. For example, in certain embodiments as described above, methodsinclude preparing the succinylated 3-(fatty acidamido)-2-hydroxy-1-(protected hydroxy)-propane organic salt bycontacting a 3-(fatty acid amido)-2-hydroxy-1-(protectedhydroxy)-propane with succinic anhydride in the presence of an organicbase. In these embodiments, methods may include contacting the crudecomposition of succinylated 3-(fatty acid amido)-2-hydroxy-1-(protectedhydroxy)-propane organic salt with the metal base directly (i.e.,without any purification or workup) to produce the succinylated 3-(fattyacid amido)-2-hydroxy-1-(protected hydroxy)-propane metal salt.

In embodiments, the fatty acid of the 3-(fatty acidamido)-2-hydroxy-1-(protected hydroxy)-propane compounds (i.e., organicsalt, metal salt, etc.) as described herein may vary, where in certaininstances, the fatty acid is selected from palmitic acid, stearic acid,oleic acid, linoleic acid, myristoleic acid and batyl fatty acid.

The protected hydroxy group of the 3-(fatty acidamido)-2-hydroxy-1-(protected hydroxy)-propane compounds as describedherein may also vary, where in certain instances, the hydroxylprotective group includes, but is not limited to: 1) an alkyl ether-typeprotective group, such as an alkyl ether, allyl ether, triphenylmethylether, dimethoxy-triphenylmethyl ether, benzyl ether or p-methoxybenzylether protecting group; 2) an ester and carbonate-type protective group,such as an acetate, chloroacetate, dichloroacetate, trichloroacetate,trifluoroacetate, pivaloate, benzoate, p-methoxybenzoate,p-bromobenzoate, methyl carbonate, 9-(fluorenylmethyl) carbonate (Fmoc),allyl carbonate (Alloc), 2,2,2-trichloroethyl carbonate (Troc),2-(trimethylsilyl)ethyl carbonate (Teoc), benzyl carbonate (Cbz),t-butyl carbonate (Boc) or dimethylthiocarbmate (DMTC) protecting group;3) an acetal type protective group, such as a methoxymethyl ether (MOM),benzyloxymethyl ether (BOM), 2,2,2-trichloroethoxymethyl ether,2-methoxymethyl ether (MEM), methylthiomethyl ether (MTM),p-methoxybenzyloxymethyl ether (PMBM), 2-(trimethylsilyl)ethoxymethylether (SEM), tetrahydropyranyl ether (THP) protecting group; and 2) asilyl ether type protective group, such as a trimethylsilyl (TMS),triethylsilyl (TES), isopropyldimethylsilyl (IPDMS),diethylisopropylsilyl (DEIPS), t-butyldimethylsilyl (TBS),t-butyldiphenylsilyl (TBDPS), triisopropylsilyl (TIPS),tetraisopropyldisiloxanylidene (TIPDS) or di-t-butylsilylene (DTBS)protecting group. In certain embodiments, the protected hydroxy group ofthe 3-(fatty acid amido)-2-hydroxy-1-(protected hydroxy)-propanecompounds described herein is a dimethoxy-triphenylmethyl protectinggroup.

In certain embodiments, the succinylated 3-(fatty acidamido)-2-hydroxy-1-(protected hydroxy)-propane metal salt is a compoundof Formula I:

In some embodiments, the present disclosure also provides an amorphoussolid succinylated 3-(fatty acid amido)-2-hydroxy-1-(protectedhydroxy)-propane lithium salt of Formula I having a peak at about 19.5°2θ in an X-ray powder diffraction pattern (XRPD, FIG. 1 ) with Cu Kαradiation. In other embodiments, the present disclosure provides anamorphous solid succinylated 3-(fatty acid amido)-2-hydroxy-1-(protectedhydroxy)-propane lithium salt of Formula I where thermogravimetricanalysis (TGA, FIG. 3 ) exhibits a single weight loss step. In theseembodiments, the single weight loss step may begin at about 225° C. Thesubject amorphous solid succinylated 3-(fatty acidamido)-2-hydroxy-1-(protected hydroxy)-propane lithium salt of Formula Imay also be characterized by TGA thermogram that shows a mass loss ofless than 1% of the total mass of the sample upon heating from 30° C. to300° C. In other embodiments, the present disclosure provides anamorphous solid succinylated 3-(fatty acid amido)-2-hydroxy-1-(protectedhydroxy)-propane lithium salt of Formula I, where the differentialscanning calorimetry (DSC, FIG. 4 ) curve exhibits an absorption peak atabout 44.9° C.

EXAMPLES

The following examples are put forth so as to provide those of ordinaryskill in the art with a complete disclosure and description of how tomake and use the present invention, and are not intended to limit thescope of what the inventors regard as their invention nor are theyintended to represent that the experiments below are all or the onlyexperiments performed. Efforts have been made to ensure accuracy withrespect to numbers used (e.g. amounts, temperature, etc.) but someexperimental errors and deviations should be accounted for. Unlessindicated otherwise, parts are parts by weight, molecular weight isweight average molecular weight, temperature is in degrees Celsius, andpressure is at or near atmospheric. By “average” is meant the arithmeticmean. Standard abbreviations may be used, e.g., bp, base pair(s); kb,kilobase(s); pl, picoliter(s); s or see, second(s); min, minute(s); h orhr, hour(s); aa, amino acid(s); kb, kilobase(s); bp, base pair(s); nt,nucleotide(s); i.m., intramuscular(ly); i.p., intraperitoneal(ly); s.c.,subcutaneous(ly); and the like.

General Synthetic Procedures

Many general references providing commonly known chemical syntheticschemes and conditions useful for synthesizing the disclosed compoundsare available (see, e.g., Smith and March, March's Advanced OrganicChemistry: Reactions, Mechanisms, and Structure, Fifth Edition,Wiley-Interscience, 2001; or Vogel, A Textbook of Practical OrganicChemistry, Including Qualitative Organic Analysis, Fourth Edition, NewYork: Longman, 1978).

Compounds as described herein can be purified by any purificationprotocol known in the art, including chromatography, such as HPLC,preparative thin layer chromatography, flash column chromatography andion exchange chromatography. Any suitable stationary phase can be used,including normal and reversed phases as well as ionic resins. In certainembodiments, the disclosed compounds are purified via silica gel and/oralumina chromatography. See, e.g., Introduction to Modern LiquidChromatography, 2nd Edition, ed. L. R. Snyder and J. J. Kirkland, JohnWiley and Sons, 1979; and Thin Layer Chromatography, ed E. Stahl,Springer-Verlag, New York, 1969.

During any of the processes for preparation of the subject compounds, itmay be necessary and/or desirable to protect sensitive or reactivegroups on any of the molecules concerned. This may be achieved by meansof conventional protecting groups as described in standard works, suchas J. F. W. McOmie, “Protective Groups in Organic Chemistry”, PlenumPress, London and New York 1973, in T. W. Greene and P. G. M. Wuts,“Protective Groups in Organic Synthesis”, Third edition, Wiley, New York1999, in “The Peptides”; Volume 3 (editors: E. Gross and J. Meienhofer),Academic Press, London and New York 1981, in “Methoden der organischenChemie”, Houben-Weyl, 4^(th) edition, Vol. 15/1, Georg Thieme Verlag,Stuttgart 1974, in H.-D. Jakubke and H. Jescheit, “Aminosauren, Peptide,Proteine”, Verlag Chemie, Weinheim, Deerfield Beach, and Basel 1982,and/or in Jochen Lehmann, “Chemie der Kohlenhydrate: Monosaccharide andDerivate”, Georg Thieme Verlag, Stuttgart 1974. The protecting groupsmay be removed at a convenient subsequent stage using methods known fromthe art.

The subject compounds can be synthesized via a variety of differentsynthetic routes using commercially available starting materials and/orstarting materials prepared by conventional synthetic methods. A varietyof examples of synthetic routes that can be used to synthesize thecompounds disclosed herein are described in the schemes below.

Example 1—Preparation of an Amorphous Solid Succinylated 3-(palmitoylamido)-2-hydroxy-1-(dimethoxytriphenylmethyl hydroxy)-propane lithiumsalt from 3-(palmitoyl amido)-2-hydroxy-1-(dimethoxytriphenylmethylprotected hydroxy)-propane (Scheme II)

3-(palmitoyl amido)-2-hydroxy-1-(dimethoxytriphenylmethyl protectedhydroxy)-propane (A) was contacted with succinic anhydride in thepresence of triethylamine in dichloromethane. The solution was stirredat room temperature for 3 hours to form the succinylated 3-(palmitoylamido)-2-hydroxy-1-(dimethoxytriphenylmethyl hydroxy)-propanetriethylamine salt intermediate (B) without isolation. The succinylated3-(palmitoyl amido)-2-hydroxy-1-(dimethoxytriphenylmethylhydroxy)-propane triethylamine salt intermediate was diluted with ethylacetate and cooled to 0-5° C. followed by addition of aqueous LiOH andstirred at 0-5° C. for 5-10 minutes. The aqueous layer and organic layercontaining the lithium salt were separated and the organic layer waswashed with LiCl. The organic layer was again separated and concentratedby rotoevaporation. The concentrate was diluted with ethyl acetate andpoured through a sintered glass funnel. The clear filtrate wasconcentrated by rotoevaporation and dried by vacuum pump until a foamedsolid was formed. Heptane was added to the foamed solid and heated to50° C. until clear. The clear solution was cooled to 45° C. over 1 houruntil hazy. Further cooling to 20° C. over 1 hour resulted in theformation of a gummy solid. The composition was further cooled to 5-10°C. until the composition formed hard solids. The slurry containing thehard solids was stirred at 10° C. overnight. The hard solids wereisolated by filtration and was dried under vacuum with a nitrogenblanket at room temperature for 30 minutes. The solid was further driedunder vacuum at ambient temperature for 16 hours.

Example 2—Preparation of an Amorphous Solid Succinylated 3-(fatty acidamido)-2-hydroxy-1-(protected hydroxy)-propane lithium salt—NormalAddition Mode

Succinylated 3-(palmitoyl amido)-2-hydroxy-1-(dimethoxytriphenylmethylprotected hydroxy)-propane triethylamine salt was converted to lithiumsalt via non-aqueous treatment of 1N LiOtBu in hexane. The crudesuccinylated 3-(palmitoyl amido)-2-hydroxy-1-(dimethoxytriphenylmethylprotected hydroxy)-propane organic salt composition was cooled to 0-5°C. followed by addition of 1N LiOtBu in hexane (1.15 equivalents) andstirred at 0-5° C. for 5-10 minutes. Heptane was added to thecomposition and concentrated by rotoevaparation down to 10-15 mL. Theaddition of heptane and concentration by rotoevaporation was repeatedtwice. The lithium salt composition in heptane was heated to 50° C.until clear. The heated composition was maintained at 50° C. for 10-15minutes under agitation, followed by cooling to 0° C. over 1 hour untilthe formation of gummy semi-solid. The slurry was aged at 0° C. for 3hours followed by filtration at 0° C. under nitrogen gas. The solidswere further dried under nitrogen and partial vacuum at 0° C. for 30minutes and then at 10° C. for 30 minutes. The solid succinylated3-(palmitoyl amido)-2-hydroxy-1-(dimethoxytriphenylmethyl protectedhydroxy)-propane lithium salt was dried under vacuum at room temperaturefor 16 hours.

Dry free-flowing solids were obtained in 91% isolated yield(uncorrected), with >99.0A % purity by HPLC, which were proved as thesuccinylated 3-(palmitoyl amido)-2-hydroxy-1-(dimethoxytriphenylmethylprotected hydroxy)-propane lithium salt by ¹H NMR (DMSO-d6).

Example 3—Preparation of an Amorphous Solid Succinylated 3-(palmitoylamido)-2-hydroxy-1-(dimethoxytriphenylmethyl hydroxy)-propane lithiumsalt—Inverse Addition Mode

To avoid the phase transition going through an oily/sticky phase,inverse addition of hot T375-Li/heptane solution into cold heptane wasdesigned and tested. The experiment was conducted in 100 mL EasyMaxvessel with intensive mixing. The hot (50° C.) heptane solution ofsuccinylated 3-(palmitoyl amido)-2-hydroxy-1-(dimethoxytriphenylmethylhydroxy)-propane lithium salt described in Example 1 was added slowly(portion-wise) to cold heptane (−10° C.). Succinylated 3-(palmitoylamido)-2-hydroxy-1-(dimethoxytriphenylmethyl hydroxy)-propane lithiumsalt solids were precipitated out immediately as uniform fine solids.The slurry formed in the heptane was aged at 0° C. for 3 hours. Aheating/cooling cycle was conducted where the cooled slurry was heatedfrom 0° C. to 20° C. over 40 minutes and maintained at 20° C. for 10minutes followed by cooling to 0° C. over 1 hour and maintained at 0° C.for 1 hour. The heating/cooling cycle was repeated twice, followed byaging of the slurry at 0° C. for 16 hours. While the slurry batchtemperature was kept at 0-10° C. there were no oily or sticky materialsobserved during and after addition of the succinylated 3-(palmitoylamido)-2-hydroxy-1-(dimethoxytriphenylmethyl hydroxy)-propane lithiumsalt solution.

The fast filtration rate of final succinylated 3-(palmitoylamido)-2-hydroxy-1-(dimethoxytriphenylmethyl hydroxy)-propane lithiumsalt solids indicated particle size increase by heat-cool cycling.De-liquor was conducted with vacuum at ambient temperature undernitrogen protection. The wet cake was dried overnight in a flask undervacuum at ambient temperature. The isolated succinylated 3-(palmitoylamido)-2-hydroxy-1-(dimethoxytriphenylmethyl hydroxy)-propane lithiumsalt product (4.92 g) gave 91% yield (uncorrected). Product loss inmother liquor was 4.6%, by HPLC analysis It should be noticed that therewas no impurity rejection by mother liquor.

The isolated dry solids of the succinylated 3-(palmitoylamido)-2-hydroxy-1-(dimethoxytriphenylmethyl hydroxy)-propane lithiumsalt from inverse addition protocol was characterized by X-ray powderdiffraction (XRPD, FIG. 1 ), polarized light microscopy (PLM, FIG. 2 ),thermogravimetric analysis (TGA, FIG. 3 ), differential scanningcalorimetry (DSC, FIG. 4 ). Lithium content of 1.36 wt % was found vs0.94 wt % in theoretical by elemental analysis.

Example 4—Physical Stability of Amorphous Solid Succinylated3-(palmitoyl amido)-2-hydroxy-1-(dimethoxytriphenylmethylhydroxy)-propane lithium salt

The physical stability of the amorphous 3-(palmitoylamido)-2-hydroxy-1-(dimethoxytriphenylmethyl hydroxy)-propane lithiumsalts were tested at room temperature under different relative humidity(RH). 300 mg of amorphous 3-(palmitoylamido)-2-hydroxy-1-(dimethoxytriphenylmethyl hydroxy)-propane lithiumsalt was placed in uncapped scintillation vials at 11% RH, 33% RH and75% RH for days. (Table 1)

TABLE 1 Physical stability of amorphous 3-(palmitoyl amido)-2-hydroxy-1-(dimethoxytriphenylmethyl hydroxy)-propane lithium salts atroom temperature under different RH RH (20-25° C.) Observation 11%Remains free-flowing solids up to 14 days* 33% 75% Turns to sticky after30 min *The samples in vials were examined by gently shaking brieflyonce in every dayAs shown in Table 1, the amorphous 3-(palmitoylamido)-2-hydroxy-1-(dimethoxytriphenylmethyl hydroxy)-propane lithiumsalts remained solid for an extended period of time at room temperatureup to 33% relative humidity. The lithium salts are stable solids thatare readily isolated and stored as compared to 3-(palmitoylamido)-2-hydroxy-1-(dimethoxytriphenylmethyl hydroxy)-propane organicsalts which formed viscous, sticky liquids at room temperature. Thelithium salts are also highly soluble in most organic solvents at orbelow ambient temperature. The high stability, ready storage and highsolubility make the 3-(palmitoylamido)-2-hydroxy-1-(dimethoxytriphenylmethyl hydroxy)-propane lithiumsalt a better starting material for use in the synthesis of compoundsthat include a fatty acid amide linker (e.g., imetelstat) as compared to3-(palmitoyl amido)-2-hydroxy-1-(dimethoxytriphenylmethylhydroxy)-propane organic salts.

What is claimed is:
 1. A method of preparing an amorphous solid composition comprising a fatty acid metal salt, the method comprising: contacting a 3-(fatty acid amido)-2-hydroxy-1-(protected hydroxy)-propane with succinic anhydride in the presence of an organic base in a polar solvent to produce a succinylated 3-(fatty acid amido)-2-hydroxy-1-(protected hydroxy)-propane organic salt; contacting the succinylated 3-(fatty acid amido)-2-hydroxy-1-(protected hydroxy)-propane organic salt with a metal base to produce a succinylated 3-(fatty acid amido)-2-hydroxy-1-(protected hydroxy)-propane metal salt as shown in Scheme I; and precipitating the succinylated 3-(fatty acid amido)-2-hydroxy-1-(protected hydroxy)-propane metal salt to produce an amorphous solid succinylated 3-(fatty acid amido)-2-hydroxy-1-(protected hydroxy)-propane metal salt composition,

wherein R is a C₁₀-C₂₀ fatty acid, ⁺Org is an organic cation, and PG is a protecting group.
 2. The method of claim 1, wherein the amorphous solid composition is a crude composition.
 3. The method of claim 2, wherein the crude composition comprises the fatty acid metal salt having a purity of 90% or more as determined by high performance liquid chromatography (HPLC), proton nuclear magnetic resonance spectroscopy (¹H NMR) or a combination thereof.
 4. The method of claim 1, wherein crude reaction products are carried through every step of the method.
 5. The method of claim 4, wherein the method is sufficient to obtain the fatty acid metal salt in an isolated yield of 90% or more as determined by high performance liquid chromatography (HPLC), proton nuclear magnetic resonance spectroscopy (¹H NMR) or a combination thereof.
 6. The method of claim 1, wherein the polar solvent comprises dichloromethane.
 7. The method of claim 1, wherein precipitating the succinylated 3-(fatty acid amido)-2-hydroxy-1-(protected hydroxy)-propane metal salt comprises: heating the succinylated 3-(fatty acid amido)-2-hydroxy-1-(protected hydroxy)-propane metal salt in a non-polar solvent to produce a heated succinylated 3-(fatty acid amido)-2-hydroxy-1-(protected hydroxy)-propane metal salt composition; and cooling the heated succinylated 3-(fatty acid amido)-2-hydroxy-1-(protected hydroxy)-propane metal salt composition to form an amorphous solid of the succinylated 3-(fatty acid amido)-2-hydroxy-1-(protected hydroxy)-propane metal salt.
 8. The method of claim 7, wherein cooling the heated succinylated 3-(fatty acid amido)-2-hydroxy-1-(protected hydroxy)-propane metal salt composition comprises inverse addition of the heated succinylated 3-(fatty acid amido)-2-hydroxy-1-(protected hydroxy)-propane metal salt composition to a cold non-polar solvent.
 9. The method of claim 7, wherein the method comprises heating the succinylated 3-(fatty acid amido)-2-hydroxy-1-(protected hydroxy)-propane metal salt in the non-polar solvent to a temperature of 45-55° C.
 10. The method of claim 7, wherein the method comprises cooling the heated succinylated 3-(fatty acid amido)-2-hydroxy-1-(protected hydroxy)-propane metal salt composition to a temperature of −10 to 0° C.
 11. The method of claim 7, wherein the non-polar solvent comprises heptane.
 12. The method of claim 7, wherein the amorphous solid of the succinylated 3-(fatty acid amido)-2-hydroxy-1-(protected hydroxy)-propane metal salt is isolated.
 13. The method of claim 1, wherein the amorphous solid of the succinylated 3-(fatty acid amido)-2-hydroxy-1-(protected hydroxy)-propane metal salt is isolated by vacuum filtration.
 14. The method of claim 1, wherein the fatty acid is selected from the group consisting of palmitic acid, stearic acid, oleic acid, linoleic acid, myristoleic acid and batyl fatty acid.
 15. The method of claim 14, wherein the fatty acid is palmitic acid.
 16. The method of claim 1, wherein the protected hydroxy group comprises a dimethoxy-triphenylmethyl protecting group.
 17. The method of claim 1, wherein the 3-(fatty acid amido)-2-hydroxy-1-(protected hydroxy)-propane is a compound of Formula II:

wherein DMT is dimethoxy-triphenylmethyl.
 18. The method of claim 1, wherein the organic salt is a triethylamine salt.
 19. The method of claim 1, wherein the metal base is selected from lithium hydroxide and lithium tert-butoxide.
 20. The method of claim 19, wherein the metal base in lithium tert-butoxide.
 21. The method of claim 1, wherein the succinylated 3-(fatty acid amido)-2-hydroxy-1-(protected hydroxy)-propane metal salt is a compound of Formula I: 